Patent Publication Number: US-2020283795-A1

Title: Plant virus movement proteins and methods of using same

Description:
RELATED APPLICATIONS 
     The present application claims priority to the U.S. Provisional Application Ser. No. 62/302,155, filed on Mar. 1, 2016, by Jessee et al., and entitled “PLANT VIRUS MOVEMENT PROTEINS AND METHODS OF USING THE SAME,” and U.S. Provisional Application Ser. No. 62/303,278, filed on Mar. 3, 2016, by Jessee et al., and entitled “PLANT VIRUS MOVEMENT PROTEINS AND METHODS OF USING THE SAME.” The entire disclosure of both of these applications is hereby incorporated by reference herein in their entirety. SEQUENCE STATEMENT 
     The Sequence Listing associated with this application is filed in electronic format via EFS-Web and is hereby incorporated by reference into the specification in its entirety. The name of the text file containing the Sequence Listing is LT01229_SL.txt. The size of the text file is 182 kilobytes, and the text file was created on Mar. 1, 2017. 
    
    
     FIELD OF THE INVENTION 
     The present invention is in the field of transfection complexes suitable for the delivery of one or more biologically active agents to a cell and methods and kits for using the same. 
     BACKGROUND OF THE DISCLOSURE 
     Lipid aggregates such as liposomes or cationic polymers can facilitate introduction of macromolecules, such as DNA, RNA, and proteins, into living cells. Aggregates comprising cationic lipid components can be used to effect delivery of large anionic molecules, such as nucleic acids, into certain types of cells. 
     The use of cationic lipids has become increasingly popular since their introduction over 25 years ago. Several cationic lipids have been described in the literature and some of these are commercially available. DOTMA (N-[1-(2,3-dioleyloxy)propyl]-N,N,N-trimethylammonium chloride; IUPAC: 1,2-di-O-octadecenyl-3-trimethyl-ammonium propane (chloride salt); CAS Number: 104872-42-6) was the first cationic lipid to be synthesized for the purpose of nucleic acid transfection. DOTMA can be formulated alone or can be combined with DOPE (dioleoylphosphatidylethanolamine) into a liposome, and such liposomes can be used to deliver plasmids into some cells. Other classes of lipids subsequently have been synthesized by various groups. For example, DOGS (5-carboxyspermylglycinedioctadecylamide) was the first polycationic lipid to be prepared and other polycationic lipids have since been prepared. The lipid DOSPA (2,3-dioleyloxy-N-[2(spermine-carboxamido)ethyl]-N,N-dimethyl-1-propanaminium) has been described as an effective delivery agent. 
     In other examples, cholesterol-based cationic lipids, such as DC-Chol (N,N-dimethyl-N-ethylcarboxamidocholesterol) have been prepared and used for transfection. Also, 1,4-bis(3-N-oleylamino-propyl)piperazine was prepared and combined with histone H1 to generate a delivery reagent that was reported to be less toxic than other reagents. Some examples of commercially available lipids include Lipofectin® (DOTMA:DOPE) (Thermo-Fisher, Carlsbad, Calif.), LipofectAmine™ (DOSPA:DOPE) (Thermo-Fisher), LipofectAmine2000™ (Thermo-Fisher), Lipofectamine 3000 (Therom-Fisher), Fugene®, Transfectam® (DOGS), ViaFect (Promega), DNA-In, GeneIn (MTI-GlobalStem) Effectene®, and DC-Chol. Further examples are PEI polymers and dendrimers such as jetPEI (PolyPlus), and Superfect (Qiagen). None of these reagents can be used universally for all cells. This is perhaps not surprising in light of the variation in composition of the membranes of different types of cells as well as the barriers that can restrict entry of extracellular material into cells. Moreover, the mechanism by which cationic lipids deliver nucleic acids into cells is not clearly understood. The reagents are less efficient than viral delivery methods and are toxic to cells, although the degree of toxicity varies from reagent to reagent. 
     However, transfection agents, including cationic lipids, anionic lipids, cationic polymers, exosomes, and virasomes, are not universally effective in all cell types. Effectiveness of transfection of different cells depends on the particular transfection agent composition. In general, polycationic lipids are more efficient than monocationic lipids in transfecting eukaryotic cells. In many cases, cationic lipids alone are not effective or are only partially effective for transfection so helper lipids or transfection enhancers can be used in combination with cationic lipids. 
     Many biological materials are taken up by cells via receptor-mediated endocytosis, in which a surface ligand binds to a cell-surface receptor, leading to clustering of ligand-bound receptors, and formation of coated pits followed by internalization of the ligands into endosomes. Both enveloped viruses, like influenza virus and alphaviruses, and non-enveloped viruses, like Adenovirus, infect cells via endocytotic mechanisms. Enhancement of dendrimer-mediated transfection of some cells by chloroquine (a lysosomotropic agent) suggests that endocytosis is involved in at least some transfections. 
     Introduction of foreign DNA sequences into eukaryotic cells mediated by viral infection is generally orders of magnitude more efficient than transfection with anionic lipids, cationic lipid, PEI, peptides, or dendrimer transfection agents. Viral infection of all the cells in a culture requires fewer than 10 virus particles per cell. Although the detailed mechanism of fusion is not fully understood and varies among viruses, viral fusion typically involves specific fusogenic agents, such as viral proteins, viral spike glycoproteins and peptides of viral spike glycoproteins. Cell binding and internalization also can be enhanced, accelerated or made selective with peptides that bind cell receptors. For example, the penton-base protein of the Adenovirus coat contains the peptide motif RGD (Arg-Gly-Asp) which mediates virus binding to integrins and viral internalization via receptor-mediated endocytosis. 
     The efficiency of cationic lipid transfections has been shown to be enhanced by the addition of whole virus particles to the transfection mixture. Certain viral components may also enhance the efficiency of cationic lipid-mediated transfection. For example, it has been suggested that “Lipofectin™”-mediated transfections may be enhanced 3-4-fold by adding influenza virus hemagglutinin peptides to the transfection mixture. Antibodies have been shown to enhance cationic lipid transfections and transferrin-poly lysine or asialoglycoprotein polylysine have been shown to enhance cationic lipid transfection. 
     Nevertheless, these methods do not work for all cell types, require relatively complex protocols and are inconvenient. It is apparent, therefore, that new and improved methods for introducing macromolecules, and particularly nucleic acids, into cell, are greatly to be desired. In particular, improved methods for introducing nucleic acids into a wider variety of cells, and particularly into primary cells, are greatly to be desired. 
     SUMMARY OF THE INVENTION 
     Disclosed herein are transfection complexes comprising at least one cell surface ligand or a plant virus movement protein or peptide fragments; at least one helper lipid component; and a transfection enhancer. Also disclosed are pharmaceutical compositions comprising the disclosed transfection complexes, and a pharmaceutically acceptable carrier. Further, disclosed are methods of transfecting a cell, the method comprising the steps of: obtaining a transfection complex as disclosed; and contacting a cell with the transfection complex. 
    
    
     DETAILED DESCRIPTION OF THE EMBODIMENTS 
     Definitions and Abbreviations 
     It is to be understood that the present invention is not limited to particular devices or biological systems, which may, of course, vary. It is also to be understood that, as used in this specification and the appended claims, the singular forms “a”, “an”, and “the” include singular and plural referents unless the content clearly dictates otherwise. Thus, for example, reference to “a lipid” includes one or more lipids. It is to be yet further understood that any terminology used herein is for the purpose of describing particular embodiments only, and is not intended to be limiting. 
     Unless stated otherwise, the following terms, definitions, and abbreviations as used herein are intended to have the following meanings: 
     As used herein, the term “labeled” is intended to mean that a compound has at least one element, isotope, or chemical compound attached to enable the detection of the compound by using a radioactive or heavy isotope label, or an immune label such as an antibody or antigen or a label derived from a colored, luminescent, phosphorescent, or fluorescent dye. Photoaffinity labeling employing, for example, o-, m- and p-azidobenzoyls, substituted with one or more halogen moieties, including, but not limited to 4-azido-2,3,5,6-tetrafluorobenzoic acid, is utilized for the direct elucidation of intermolecular interactions in biological systems. 
     The terms “subject” and “animal” are synonymous and, as used herein, refer to humans as well as non-human animals, including, for example, mammals (e.g., a rodent, a mouse, a rat, a rabbit, a monkey, a dog, a cat, a primate, or a pig), birds, reptiles, amphibians, and fish. 
     The term “cell” generally refers to eukaryotic cells of any type and from any source. Types of eukaryotic cells include epithelial, fibroblastic, neuronal, hematopoietic cells and the like from primary cells, tumor cells or immortalized cell lines. Sources of such cells include any animal such as human, canine, mouse, hamster, cat, bovine, porcine, monkey, ape, sheep, fish, insect, fungus, and any plant including crop plants, algae, ornamentals and trees. 
     “Delivery” is used to denote a process by which a desired compound is transferred to a target cell such that the desired compound is ultimately located inside the target cell or in, or on, the target cell membrane. In many uses of the compounds of the invention, the desired compound is not readily taken up by the target cell and delivery via lipid aggregates or transfection complexes a means for delivering the desired compound to the appropriate cellular compartment within a cell. In certain uses, especially under in vivo conditions, delivery to a specific target cell type is preferable and can be facilitated by transfection complexes comprising surface ligands of the invention. 
     Drug refers to any therapeutic or prophylactic agent other than food which is used in the prevention, diagnosis, alleviation, treatment, or cure of disease in man or animal. 
     “Kit” refers to transfection or protein expression kits which include one or more of the compounds of the present invention or mixtures thereof. Such kits may comprise a carrying means being compartmentalized to receive in close confinement one or more container means such as vials, test tubes and the like. Each of such container means comprises components or a mixture of components needed to perform transfection. Such kits may include one or more components selected from nucleic acids (preferably one or more vectors), cells, one or more compounds of the present invention, lipid-aggregate forming compounds, transfection enhancers, biologically active substances, etc. 
     The term “associated with”, when used in the context of molecular interactions, refers to two entities linked by a direct or indirect covalent or non-covalent interaction, such as hydrogen bonding, van der Waals interactions, hydrophobic interactions, magnetic interactions, electrostatic interactions, etc. 
     The term “biocompatible,” as used herein refers to compounds that are not toxic to cells. Compounds are biocompatible if their addition to cells in vitro results in less than or equal to 20% cell death, and their administration in vivo does not induce inflammation or other such adverse effects. 
     The term “biodegradable,” as used herein, refers to compounds that, when introduced into cells, are broken down into components that the cells can either reuse or dispose of without significant toxic effect on the cells (i.e., fewer than about 20% of the cells are killed when the components are added to cells in vitro). The components do not induce inflammation or other adverse effects in vivo. The chemical reactions relied upon to break down the biodegradable compounds are typically uncatalyzed. The term “effective amount,” as used herein with respect to an active agent, refers to the amount necessary to elicit the desired biological response. The effective amount of an agent or device may vary depending on such factors as the desired biological endpoint, the agent to be delivered, the composition of the encapsulating matrix, the target tissue, etc. Delivery of an “effective amount of a molecule” is the delivery of the molecule into a cell in sufficient amounts so that the molecule elicits a biological response, for example, modulating the expression of one or more genes in the cell. In specific embodiments, an effective amount of a molecule is delivered to a cell such that an amelioration or improvement in a disease, condition, or disorder related to the cell can be obtained. Delivery of an “effective amount of siRNA” or an “effective amount or RNAi” is the delivery of siRNA or other RNAi into a cell in sufficient amounts to cause a reduction in expression of the target gene in the cell. 
     The terms “biologically active agent”, “bioactive agents” or the like, generally refers to a composition, complex, compound or molecule which has a biological effect or that modifies, causes, promotes, enhances, blocks or reduces a biological effect, or that enhances or limits the production or activity of, reacts with and/or binds to a second molecules which has a biological effect. The second molecule can, but need not be, an endogenous molecule (e.g., a molecule, such as a protein or nucleic acid, normally present in the target cell). A biological effect may be, but is not limited to, one that stimulates or causes an immunoreactive response; one that impacts a biological process in a cell, tissue or organism (e.g., in an animal); one that imparts a biological process in a pathogen or parasite; one that generates or causes to generate a detectable signal; one that regulates the expression of a protein or polypeptide; one that stops or inhibits the expression of a protein or polypeptide; or one that causes or enhances the expression of a protein or polypeptide. Biologically active compositions, complexes, compounds or molecules may be used in investigative, therapeutic, prophylactic and diagnostic methods and compositions. Bioactive agents may include but are not limited to, pharmaceuticals, cell metabolites, proteins, nutrients (vitamins, amino acids, lipids, nucleotides, and carbohydrates), and exosomes. 
     The term “cationic lipid” refers to any cationic lipids which may be used for transfection and which under physiological conditions possess at least one positive charge. While it is to be understood that certain of the cell surface ligands that form the basis of the present disclosure can be formulated with cationic lipids the cationic lipids can be considered helper lipids. 
     The term “lysosomotropic agent” is any compound or molecule which inhibit lysosomal function that prevents or slows the acidification of the lysosomal compartment. 
     The term “nucleic acid binding moiety” as used herein refers to a compound or molecule capable binding to nucleic acid. In some embodiments, the binding molecule is capable of noncovalently binding to nucleic acid, while in other embodiments, the binding molecule links covalently to a cell binding adhesion sequence, a plant virus movement protein or peptide fragments, a nuclear localization sequence, transfection enhancer, and/or a fusion agent. The binding molecule can include but is not limited to spermine, spermine derivative, spermidine, histones or fragments thereof, protamines or fragments thereof, HMG proteins or fragments thereof, poly-lysine, poly-arginine, poly-histidine, polyamines and cationic peptides, nucleic acid intercalaters, protein nucleic acid sequences or aptamers. In addition, this includes but is not limited to analogs or derivatives of the above compounds. Non limiting examples are the cationic peptides that are repeats of lysine or arginine, for example a sequence having between 8-20 lysine residues (K8-K20) (SEQ ID NO:583) or between 8-20 arginine residues (R8-R20) (SEQ ID NO:584). 
     “Target cell” or “target tissue” refers to any cell or tissue to which a desired compound is delivered, using a lipid aggregate or transfection complex as carrier for the desired compound. 
     Transfection is used herein to mean the delivery of any nucleic acid, protein, peptide, lipid, cell nutrient, pharmaceutical agent, molecule or other macromolecule to a target cell or tissue in vitro or in vivo (i.e., in an animal, a plant or a human), such that the nucleic acid, protein or other macromolecule is expressed in, confers a phenotype to, causes enhanced growth, expression of a protein, or has a biological function in the cell. 
     The term “expressible nucleic acid” includes both DNA and RNA without regard to molecular weight, and the term “expression” means any manifestation of the functional presence of the nucleic acid within the cell including, without limitation, both transient expression and stable expression. 
     The term “fusion agent” as used herein refers to any chemical or molecules capable breaking down an endosomal membrane or cell membrane and freeing the transfection agent into the cytoplasm of the cell. This term includes but is not limited to viruses, synthetic compounds, proteins, fusion peptides, cell penetration peptides or proteins, or derivatives thereof. As a result of the presence of the fusion agent the membrane can undergo lysis, fusion, or rearrangement or all three. 
     The term “fusion peptide” refers to any peptide grouping which penetrates a membrane such that the structural organization and integrity of the membrane is lost. Fusion peptides are fusion agents. 
     The term “transfection agent” as used herein generally refers to composition capable of delivering molecules to cells. Transfection agents can be organic such as lipid, carbohydrate, cationic polymers, dendrimers, peptide or protein based or combination of those depending cell type or tissue that one targets. Transfection agents can also be in-organic such as calcium salts. They included cationic lipids, anionic lipids, cationic peptides, cationic proteins, polycationic virus hybrids, cationic polymers, exosomes, and combinations of the above. Transfection agent as used herein may optionally include at least one or more of the transfection compounds optionally in combination with one or more helper lipids, one or more pegylated lipids, one or more lipids from exosomes, complete lipid mixtures form exosomes, optionally one or more targeting moieties, optionally one or more cell surface ligands, optionally one or more nuclear localization sequences, optionally one or more fusion agents, optionally one or more condensing agents, optionally one or more cell penetration agents, optionally one or more plant movement proteins or peptide fragments, optionally one or more exosomes and optionally one or more lysosomotropic agents. 
     The term “transfection enhancer” as used herein refers to a compound when added to a transfection agent increases the efficiency of transfection (i.e., increases the percent of cells transfected), increases the level of expression of a transfection agent, or reduces the requirement for the amount of nucleic acid or protein required to give a biological response, or any combination of the enhancements above. In some embodiments, the transfection enhancer also helps deliver molecules that help downregulate expression such as siRNA, LNA&#39;s and the like. 
     The term “surface ligand” or “cell surface ligand” refers to a chemical or structure which will bind to a surface receptor of a cell. The term “cell surface receptor” as used herein refers to any specific chemical grouping on the surface of a cell to which the surface ligand can attach, contact or associate with. A surface ligand is a targeting moiety. Furthermore, surface ligands include anything which is capable of binding to the cell and centering the cell through cytosis (e.g., endocytosis, potocytosis, and pinocytosis). 
     The term “transfection complex”, as used herein generally refers to a composition formulated for the delivery of a biologically active agent, such as a nucleic acid, a protein, a macromolecule, cell nutrient, bioactive molecule or the like, to a cell or to a tissue in vivo or in vitro. Transfection complexes as used herein may include at least one or more of the transfection compounds or agents in combination with the biologically active compound to be delivered, optionally in combination with; one or more helper lipids, one or more pegylated lipids, one or more targeting moieties, one or more nuclear localization sequences, one or more fusion agents, one or more condensing agents, one or more cell penetration agents, one or one or more plant movement proteins or peptide fragments, complete exosomes, total lipid extracts isolated from exosomes, one or more exosome lipids, one more exosome protein components and one or more lysosomotropic agents in addition to the bioactive agent that is to be delivered. For the purposes described herein, the term “transfection complex” may be thought of as a lipoplex or a lipid aggregate contacted with a bioactive agent. Thus, in some instances in the following disclosure, terms such as lipoplex, lipid aggregate and the like may be used to make reference a transfection complex that lacks the one or more bioactive agents or “payloads”. 
     The term “helper lipid”, as used herein, generally refers to a lipid that is suitable for use in the preparation and formation of transfection complexes disclosed herein. Suitable helper lipids may include, though are not limited to DOPE, DPhPE, saturated and unsaturated DPPE, saturated and unsaturated DMPE, DOPC, Lyso-PE (1-acyl-2-hydroxy-sn-glycero-3-phosphoethanolamine), Lyso-PC (1-acyl-3-hydroxy-sn-glycero-3-phosphocholine), 3-alkyloxy-2-hydroxy-1-acetamidopropane, 4-alkyloxy-3-hydroxy-1-acetamidopropane, 5-alkyloxy-4-hydroxy-1-acetamidopropane, cholesterols, cholesterol derivatives, sterols, including phytosterols, zoosterols and hopanoids, or any of the neutral or cationic lipids that are known to allow or to facilitate the introduction of exogenous bioactive molecules to the interior of a cell or of a tissue. In some embodiments, more than one helper lipid may be used in the formulation of the transfection complexes described herein. Exemplary though non-limiting neutral or cationic lipids contemplated for use in the preparation of the presently disclosed transfection complexes may include one or more lipids selected from the following: N-(2-bromoethyl)-N,N-dimethyl-2,3-bis(9-octadecenyloxy)-propana minimun bromide (BMOP), dipalmitoylphosphatidylethanolamine 5-carboxyspermylamide (DDPES), DSPC, dioleoylphosphatidylethanolamine (DOPE), formulation of cetyltrimethylammonium bromide (CATB) and DOPE (CTAB:DOPE), 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphocholine (POPC), DMG, 1,2-dimyristloxyaminopropane (DMAP), dimyri stoylphospatidylethanolamine (DMPE), DOMG, DMA, Dioleoylphosphatidylcholine (DOPC), dimyristoylphosphatidylcholine (DMPC), Dipalmitoylethylphosphatidylcholine (DPEPC), dioleoydimethylammonium chloride (DODAC), 1,3-di-oleoyloxy-2-(6-carboxyspermyl)-propylamid (DOSPER), N-[1-(2,3-dioleyloxy)propyl]-N,N,N-trimethylammoniumchloride (DOTMA), N-[1-(2,3-dipalmitoleoyloxy)propyl]-N,N,N-trimethylammoniumchloride (DPTMA), didoceyl methylammonium bromide (DDAB), N-[1-(2,3-dioleoyloxy)propyl]-N,N,N-trimethyl-ammonium methylsulfate (DOTAP), DOTAP.Cl, 3,β-N,(N′,N′-dimethylaminoethane)-carbamoyl]cholesterol (DC-chol), 2-(sperminecarboxamido)ethyl)-N,N-dimethyl-lammonium trifluoroacetate (DOSPA), 0,0′-ditetradecanoyl-N-(alphatrimethylammonioacetyl) diethanolamine chloride (DC-6-14), dicaproylphosphtidylethanolamine (DCPE), dilauryl oxypropyl-3-dimethylhydroxy ethylammonium bromide (DLRIE), 1,2-dioleoyl-3-dimethylammonium-propane (DODAP), Ethyl-PC, 2,3-dioleoyloxy-N-[2-(sperminecarboxamidoethyl]-N,N-di-met-hyl-1-propanaminium trifluoroacetate (DOSPA), dioctadecylamidoglycyl carboxyspermine (DOGS), N-[1-(2,3 dimyristyloxy)propyl]-N,N-dimethyl-N-(2-hydroxyethyl) ammonium bromide (DMRIE), Dioleoylethyl-phosphocholine (DOEPC), N-[1-(2,3-dioleoyloxy)propyl]-N-[1-(2-hydroxyethyl)]-N,Ndimethylammonium iodide (DOHME), N-(3-aminopropyl)-N,N-dimethyl-2,3-bis(dodecyloxy)-1-propaniminium bromide/dioleylphosphatidylethanolamine (GAP-DLRIE:DOPE), dipalmitoylphospha-tidylcholine (DPPC), 1,2-dioleoyl-sn-glycero-3-[phospho-rac-(3-lysyl(1-glycerol)).Cl (DOPG), N-lauroylsarcosine, (R)-(+)-limonene, lecithins (and derivatives thereof); phosphotidylethanolamine (and derivatives thereof); phosphatidylethanolamines, dioleoylphosphatidylethanolamine), diphytanoylphosphatidylethanolamine (DPhPE), dipalmitoylphosphatidylethanolamine (DPPE), dipalmiteoylphosphatidylethanolamine, 3-β-[1-ornithinamidecarbamoyl]-cholesterol (O-Chol), palmitoyloleoylphosphatidyl-ethanolamine (POPE); di stearoylphosphatidylethanolamine; phosphotidylcholine; phosphatidylcholines, dipalmitoylphosphatidylcholine (DPPC) palmitoyloleoyl-phosphatidylcholine (POPC); distearoylphosphatidylcholine; phosphatidylglycerol; piperazine-based cationic lipids, a phosphatidylglycerol, dioleoylphosphatidylglycerol (DOPG), dipalmitoylphosphatidyl-glycerol (DPPG), distearoylphosphatidylglycerol; phosphatidylserine (and derivatives thereof); phosphatidylserines, dioleoyl- or dipalmitoylphosphatidylserine; a diquaternary ammonium salt; N,N′-dioleyl-N,N,N′,N′-tetramethyl-1,2-ethanediamine (TmedEce), N,N′-dioleyl-N,N,N′,N′-tetramethyl-1,3-propanediamine (PropEce), N,N′-dioleyl-N,N,N′,N′-tetramethyl-1,6-hexanediamine (HexEce), and their corresponding N,N′-dicetyl saturated analogues (TmedEce, PropEce and HexEce), a diphosphatidylglycerol; a fatty acid ester; a monocationic transfection lipid; 1-deoxy-1-[dihexadecyl(methyl)ammonio]-D-xylitol; 1-deoxy-1-[methyl(ditetra-decyl)ammonio]-Darabinitol; 1-deoxy-1-[dihexadecyl(methyl)ammonio]-D-arabinitol; a 1-deoxy-1-[methyl(dioctadecyl)-ammonio]-darabinitol, glycerol ester; sphingolipids; cardolipin; a cerebroside; a ceramide, exosomes or lipids mixtures isolated from exosomes; and combinations thereof. 
     Helper lipids also include the neutral lipids cholesterol and other 3βOH-sterols, as well as derivatives thereof, phosphatidyl choline, or commercially available cationic lipid mixtures such as, for example, LIPOFECTIN® CELLFECTIN® (1:1.5 (M/M) formulation of N,N,N′,N″,N″′-tetramethyl-N,N,N′,N″,N″′-tetrapalmitylspermine (TMTPS) and dioleoyl phosphatidylethanolamine (DOPE), LIPOFECTACE®, GS 2888 CYTOFECTIN®, FUGENE 6®, EFFECTENE®, and LIPOFECTAMINE®, LIPOFECTAMINE 2000®, LIPOFECTAMINE PLUS®, LIPOTAXI®, POLYECT®, SUPERFECT®, TFXNT™, TRANSFAST™, TRANSFECTAM®, TRANSMESSENGER®, vectamidine (3-tetradecylamino-N-tert-butyl-N′-tetradecylpropionamidine (a.k.a. diC14-amidine), OLIGOFECTAMINE MessengerMAX, GeneIn™, TransfeX™, LipofectAmine 3000, Lipofectin®, DMRIE-C, CellFectin®, LipofectAce®, Fugene®, Fugene® HD, Tfx-10®, Tfx-20®, Tfx-50®, DNA-In, Transfectin™, SilentFect™, Effectene®, ViaFect™, DC-chol, GenePorter®, DharmaFect 1®, DharmaFect 2®, DharmaFect 3®, DharmaFect 4®, Escort™ III, Escort™ IV, DOGS among others. Also contemplated are any mixtures of combination of the above listed helper lipids, exosomes, and lipids mixtures isolated from exosomes. 
     Examples of lipids examples isolated from exosomes are include, but are not limited to, Lyso-PC (non-limiting examples include C-18, C-16, C-14 and mixture), lyso-bisphospahtidic acid (non-limiting example include C-18, C-16 and C-14), sphingomyelin, ceramides (non-limiting examples include C-8 and C-24), disaturated PC (non-limiting examples include DSPC, DPPC, DMPC, and compounds having a Cn length (where n=8-25), diunsaturated PC-MIX (non-limiting examples include DOPC and DP(db)PC), phosphatidyl serine (PS), phosphatidyl inositol (PI), disaturated PE (non-limiting example include DSPE, DPPE, and DMPE), di-unsaturated PE-MIX (non-limiting example include DOPE and DP(db)PE), posphatidyl glycerol (PG), (non-limiting examples include C-18-C-22), cholesterol, and diglycerides, such as cardiolipin. 
     Also contemplated are any mixtures of combination of the above listed helper lipids, exosomes, and lipids mixtures isolated from exosomes. 
     The following patent documents, patent applications, or references are incorporated by reference herein in their entirety and in particular for their disclosure of transfection agents containing cationic and neutral helper lipids, which may be used in the transfection complexes disclosed herein: U.S. Pat. Nos. 6,075,012; 6,020,202; 5,578,475; 5,736,392; 6,051,429; 6,376,248; 5,334,761; 5,316,948; 5,674,908; 5,834,439; 6,110,916; 6,399,663; 6,716,882; 5,627,159; 7,915,230; 7,531,693; 8,034,977; 7,166,745; 5,994,109; 6,033884; 6,150,168; 6,177,554; 6,083,741 6,458,026; 7,598,421; 7,820,624; 7,256,043; 7,704,969; 8,026,341; 7,145,039; 7,531,693; and 8,785,200; and International Publications WO 2004/063342, WO 0027795, WO 2004/105697, WO 2007/130073, WO 2012/142622, and WO 2013/158127, 
     The term “pegylated lipid” as used herein generally refers to a lipid that is covalently conjugated to one or more polyethylene glycol moieties. Pegylated lipids for lipoplex embodiments herein include phosphatidylethanolamine (PE) based pegylated lipids such as, for example, 1,2-dimyristoyl-sn-glycero-3-phosphoethanolamine-N-[methoxy(polyethylene glycol)-MW] where MW refers to average MW of the polyethylene glycol moiety. Such dimyristoyl-PEG-PE lipids are commonly designated 14:0 PEG (MW) PE. The average MW of the polyethylene glycol moiety can be 25, 350, 550, 750, 1000, 2000, 3000, 5000, 6000, 8000 or 12000, for example. The fatty acid chains of the phosphatidylethanolamine based pegylated lipids may include, for example, a 1,2-dioleoyl group such as for 18:1 PEG (MW) PE, a 1,2-dipalmitoyl group such as for 16:0 PEG (MW) PE, or a 1,2-distearoyl-group such as for 18:0 PEG (MW) PE. Further phosphatidylethanolamine (PE) based pegylated lipids include, for example, 1,2-distearoyl-sn-glycero-3-phosphoethanolamine-N-[MOD(polyethylene glycol)-MW], also referred to as DSPE-MOD PEG(MW) wherein MOD refers to a functional moiety such as an amine, biotin, carboxylic acid, folate, maleimide, PDP, or carboxyfluorescein moiety. The MW may be 2000 or 5000, for example. Pegylated lipids for the embodiments described herein also include ceramide based pegylated lipids such as, for example, N-octanoyl-sphingosine-1-{succinyl[methoxy(polyethylene glycol)MW]}, designated C8 PEG (MW) ceramide, where MW is 750, 2000, or 5000, for example. Alternatively, the fatty acid moiety may have an N-palmitoyl (C16) group such as for C16 PEG (MW) ceramide. 
     A “liposomal composition” generally is a formulation that includes one or more liposomes. In some instances, the term “liposomal composition” may be used interchangeably with the term “transfection complex”. These formulations are typically colloids, but can be dried formulations as well. A liposome is a vesicular colloidal particle composed of self-assembled amphiphilic molecules. Surface ligands, plant virus movement proteins, or fragments thereof disclosed herein can be incorporated into liposomal compositions of one or more cationic lipids, or one or more anionic lipids either or one or more pH sensitive lipids alone or optionally in combination with one or more helper lipids (i.e., a neutral lipid, a cholesterol or cholesterol derivative, lysolipid. or cationic lipids) that are processed using standard methods to form a liposome-containing colloid suspension. Liposomal compositions disclosed herein are those containing one or more cationic lipids, one or more helper lipids, optionally, in combination with one or more neutral and/or helper lipids, exosomes, total lipid extract from exosomes, targeting moieties, fusion agents, cell penetration agents, lysomotropic agents which are treated by any of the standard methods known in the art without limitation to form liposomes. The liposomal compositions may optionally contain one or more fusion agents. The liposomal compositions may optionally contain one or more liposomal compositions can be distinguished one from another by particle size measurements. Different compositions will exhibit differences in particle size and uniformity of particle size, e.g., average particle size, and polydispersity. Different compositions will exhibit differences in the extent of the composition that is in the form of liposomes. In some non-limiting embodiments, liposomal compositions will exhibit particle size in the range 120 nm and 800 nm and will exhibit generally lower polydispersity. Lipoplex particle size (with siRNA or other cargo) may range from about 40 nm to 135 nm. In some embodiments, lipoplex particle size is 50 nm to 120 nm, 50 nm to 100 nm, 60 nm to 90 nm, 70 nm to 90 nm, or about 85 nm. 
     The term “Lipid aggregate” or “lipoplex” is a generic term that includes liposomes of all types, both unilamellar and multilamellar, as well as vesicles, micelles, exosomes, micro-vesicles and more amorphous aggregates. A cationic lipid aggregate is a lipid aggregate comprising a combination of one or more cationic compounds, optionally in combination with non-cationic lipids (including neutral lipids), exosomes, such that the lipid aggregate has a net positive charge. Surface ligands or plant virus movement proteins or fragments thereof disclosed herein can be incorporated into lipid aggregate, optionally with a helper lipid and further optionally with one or more pegylated lipids and/or one or more targeting moieties, one or more fusion agents, one or more cell penetration agents and one or more lysosomotropic agents, one or more exosomes, which can then form a lipid-bioactive agent complex when contacted with a suitable bioactive agent. The terms “lipid aggregate” or “lipoplex” are generally used herein to refer to a “naked” transfection complex, i.e., a transfection complex that generally lacks a payload of bioactive agent to be delivered to a cell or to a tissue in vitro or in vivo. 
     The term “exosome” refers to the small membrane vesicles secreted by most cells that contain cell specific payloads of proteins, lipids and, genetic material and other biomolecules that are transported to other cells in different location of the tissue. Exosomes can be considered liposomal particles. Exosomes or lipid mixtures obtained therefrom, can be used in combination with other transfection agents or helper lipid mixtures. Exosomes are also referred to as microvesicles, epididimosomes, argosomes, exosome-like vesicles, microparticles, promininosomes, prostasomes, dexosomes, texosomes, archeosomes and oncosomes In one example of lipid constituents of exosomes is Lyso-PC (non limiting examples of which C-18, C-16, C-14 and mixture), Lyso-bisphospahtidic acid (non limiting example of which is C-18, C-16 and C-14), Sphingomyelin, Ceramides (non limiting examples C-8-C-24), Disaturated PC (non limiting examples (DSPC, DPPC, DMPC and others where Cn (n=8-25) Diunsaturated PC-MIX (non limiting examples of which are DOPC, DP(db)PC) phosphatidyl serine (PS), phosphatidyl inositol (PI), Disaturated PE (non limiting example, DSPE, DPPE, DMPE), Di-unsaturated PE-MIX (non limiting example DOPE DP(db)PE), posphatidyl glycerol (PG), (non limiting examples of which are C-18-C-22, Cholesterol, Diglycerides such as cardiolipin 
     The term “lipid-bioactive agent” generally refers to the noncovalent association between a lipid or lipid aggregate and a bioactive agent, such as a nucleic acid, nucleotide, amino acid, peptide, a polypeptide, protein, protein nucleic complex, nutrient, exosome and the like. 
     As used herein “nucleic acid” and its grammatical equivalents will include the full range of polymers of single or double stranded nucleotides and includes nucleic acids (including DNA, RNA, and DNA-RNA hybrid molecules, Linked Nucleic acids (LNA), Bridged Nuclic acid (BNA)) that are isolated from a natural source; that are prepared in vitro, using techniques such as PCR amplification or chemical synthesis; that are prepared in vivo, e.g., via recombinant DNA technology; or that are prepared or obtained by any known method. A nucleic acid typically refers to a polynucleotide molecule comprised of a linear strand of two or more nucleotides (deoxyribonucleotides and/or ribonucleotides) or variants, derivatives and/or analogs thereof. The exact size will depend on many factors, which in turn depends on the ultimate conditions of use, as is well known in the art. The nucleic acids of the present invention include without limitation primers, probes, oligonucleotides, vectors, constructs, plasmids, genes, transgenes, genomic DNA, cDNA, LNA, BNA, RNA, mRNA, tRNA, miRNA, RNAi, siRNA, shRNA, stRNA, guide-RNA, gBlock, PCR products, restriction fragments, oligonucleotides and the like. 
     As used herein, the term “nucleotide” includes any monomeric unit of DNA or RNA containing a sugar moiety (pentose), a phosphate, and a nitrogenous heterocyclic base and may also include mono-, di- and triphosphate forms of such nucleotides. The base is usually linked to the sugar moiety via the glycosidic carbon (at the 1′ carbon of pentose) and that combination of base and sugar is called a “nucleoside.” The base characterizes the nucleotide with the four customary bases of DNA being adenine (A), guanine (G), cytosine (C) and thymine (T). Inosine (I) is an example of a synthetic base that can be used to substitute for any of the four, naturally occurring bases (A, C, G, or T). The four RNA bases are A, G, C, and uracil (U). Accordingly, a nucleic acid may be a nucleotide sequence comprising a linear array of nucleotides connected by phosphodiester bonds between the 3′ and 5′ carbons of adjacent pentoses. Other modified nucleotides are known and may be sued in the practice of the invention. The term nucleotide includes ribonucleoside triphosphates ATP, UTP, ITP, CTG, GTP or derivatives such as but not limited to [aS] ATP, 7-deaza-GTP and 7-deaza-ATP, 5-methyCTP, pseudoUTP, 4-thioUTP and deoxyribonucleoside triphosphates such as dATP, dCTP, dITP, dUTP, dGTP, dTTP, or derivatives thereof. Such derivatives include, for example, [αS]dATP, 7-deaza-dGTP and 7-deaza-dATP, 5-methydCTP, pseudodUTP, 4-thiodUTP, LNA-Nucleosidetriphosphates and nucleotide derivatives that confer nuclease resistance on the nucleic acid molecule containing them. The term nucleotide as used herein also refers to dideoxyribonucleoside triphosphates (ddNTPs) and their derivatives. Illustrated examples of dideoxyribonucleoside triphosphates include, but are not limited to, ddATP, ddCTP, ddGTP, ddITP, and ddTTP. The term nucleotides as used here also refer to nucleotides that contain modifiable groups. Illustrated examples of nucleotides with modifiable group include, but are not limited to, allyamine-CTP, allyamine dCTP, allyamine UTP, allyamine dUTP. According to the present invention, a “nucleotide” may be unlabeled or detectably labeled by well-known techniques. Detectable labels include, for example, biotin, radioactive isotopes, fluorescent labels, chemiluminescent labels, bioluminescent labels and enzyme labels. Various labeling methods known in the art can be employed in the practice of this invention. Transfection complexes of this invention can be used to deliver nucleotides to living cells to allow incorporation of modified nucleotides in nucleic acids. 
     “RNA” or “RNA molecule” refers to any RNA molecule or functional portion thereof, of any size, self-replicating, and having any sequence, from any source, including RNA viral, prokaryotic, and eukaryotic organisms. The RNA molecule may be chemically modified and in any form, including, but not limited to, linear or circular, and single or double stranded. Non-limiting examples of RNA molecules include mRNA, rRNA, tRNA, miRNA, mtRNA, tmRNA, RNAi, siRNA, shRNA, guideRNA, and stRNA. In some embodiments, siRNA molecules useful in the practice of the invention include, for example, those described in U.S. Patent Publication Nos. 2004/0014956, 2004/0054155, 2006/0009409, 2009/0023216, and 2010/0136695; and as described in International Publications WO 2003/064626, and WO 03/064625, all of which are incorporated by reference herein. Further siRNA molecules useful in the practice of the invention include, for example, those described in International Publication WO 2009/039173, which application is incorporated by reference herein. 
     The terms “peptide”, “polypeptide”, or “protein,” as used herein refer to a string of at least three amino acids linked together by peptide bonds. The terms “protein” and “peptide” may be used interchangeably, though it is generally understood that a “polypeptide” or “protein” is larger than a peptide. “Peptide” may refer to an individual peptide or a collection of peptides. 
     The terms “polynucleotide” or “oligonucleotide,” as used herein, refer to a polymer of nucleotides. Typically, a polynucleotide comprises at least three nucleotides. The polymer may include natural nucleosides (i.e., adenosine, thymidine, guanosine, cytidine, uridine, deoxyadenosine, deoxythymidine, deoxyguanosine, and deoxycytidine), nucleoside analogs (e.g., 2-aminoadenosine, 2-thiothymidine, inosine, pyrrolo-pyrimidine, 3-methyl adenosine, C5-propynylcytidine, C5-propynyluridine, C5-bromouridine, C5-fluorouridine, C5-iodouridine, C5-methylcytidine, 7-deazaadenosine, 7-deazaguanosine, 8-oxoadenosine, 8-oxoguanosine, O(6)-methylguanine, and 2-thiocytidine), chemically modified bases, biologically modified bases (e.g., methylated bases), intercalated bases, modified sugars (e.g., 2′-fluororibose, ribose, 2′-deoxyribose, arabinose, and hexose), or modified phosphate groups (e.g., phosphorothioates and 5′-N-phosphoramidite linkages). The term “lipid” refers to hydrophobic or amphiphilic organic compounds inclusive of fats, oils and triglyderides. 
     DISCLOSED ASPECTS AND EMBODIMENTS 
     Transfection complexes suitable for the delivery of one or more biologically active agents to a cell or a tissue in vitro or in vivo are provided for herein. The transfection complexes described herein include one or more cell surface ligands, plant virus movement proteins, or fragments thereof that enhance transfection in combination with a transfection agent as part of a transfection complex. In some embodiments, the transfection complexes disclosed herein optionally further comprise one or more components selected from the group consisting of one or more helper lipids, one or more pegylated lipids, one or more cationic lipids, one or more cationic polymers one or more targeting moieties, exosomes, total lipid isolated from exosomes, total lipid and protein isolated form exosomes, a combination thereof. In some embodiments, transfection complexes disclosed herein further comprise one or more components selected from the group consisting of peptide or non-peptide transfection enhancers, consisting of peptide or non-peptide surface ligand, cell penetration peptide or non-peptide cell penetration agent, fusogenic peptide or non-peptide fusion agents, peptide or non-peptide endosomal release agents, lysomolotropic agents, nuclear targeting agents (such as, e.g., a peptide containing one or more nuclear localization sequences), and a combination thereof. 
     Plant virus movement proteins are involved in moving the genome of plant viruses from cell to cell and from cell membrane to nucleus and back to the cell membrane and then into the neighboring cells using the plant host cell machinery in combination with the plant virus movement protein. All plant viruses have movement protein but no all have been studied. These virus movement proteins, or fragments of such have utility for moving DNA or RNA in eukaryotic cells from the cell cytoplasium to the nucleus or nucleus to cell membrane. The virus genera and virus families of Tobamovirus, Dianthovirus, Umbravirus, Bromovirus, Cucumovirus, Begomovirus, Potyvirus, Hordei-like virus, Potex-like virus, Begomoviruses, Geminiviridae have example of virus movement proteins that move nucleic acid to and from the cell membrane and to the nucleus if a DNA virus. Utilization of these plant proteins to enhance transfection of both DNA and RNA are disclosed. 
     In some embodiments, helper lipids suitable for use in the preparation and formation of transfection complexes disclosed herein include, though are not limited to a cholesterol, a cholesterol derivative, one or more sterols, including phytosterols, zoosterols and hopanoids, or any of the neutral or cationic lipids that are known to allow or to facilitate the introduction of exogenous bioactive molecules to the interior of a cell or of a tissue. The helper lipid or helper lipids may be derived form exosomes or complete exosomes. In some embodiments, more than one helper lipid is used in the formulation of the transfection complexes described herein. In some embodiments, the transfection complexes disclosed herein comprise no helper lipid. 
     Illustrative though non-limiting neutral or cationic lipids suitable for use as helper lipids in accordance with some of the embodiments set forth herein include saturated and unsaturated alkyl and alicyclic ethers and esters of amines, amides or derivatives thereof. In some embodiments, straight-chain or branched alkyl and alkene groups of cationic lipids contain from 1 to about 25 carbon atoms. In certain embodiments, straight-chain or branched alkyl or alkene groups have six or more carbon atoms. In some embodiments, straight-chain or branched alkyl or alkene groups have eight to about twenty carbon atoms. In other embodiments, alicyclic groups contain from about 6 to 30 carbon atoms, or, alternatively, eight to twenty carbon atoms. In some embodiments, the alicyclic groups include cholesterol and other steroid groups. In certain embodiments, cationic lipids are prepared with a variety of counter ions (anions) including among others: a halide (i.e., Cl − , Br − , I − , F − ), acetate, trifluoroacetate, sulfate, nitrite, triflate, and nitrate 
     Embodiments of pegylated lipids suitable for use in the preparation and formation of the transfection complexes disclosed herein are any lipid or mixture of lipids that are compatible with the formation of transfection complexes described herein, and with the administration thereof to an animal or to a human in vivo, or to tissues or cells in vitro. The pegylated lipids used with the presently described transfection complexes include, but are not limited to, a PEG polymer having a molecular weight between about 250 daltons and about 12,000, or in some embodiments, about 350 daltons and about 6,000 daltons, or, in some embodiments, between about 500 daltons and about 1,000 daltons, or, in some embodiments, between about 1,000 daltons and about 2,000 daltons, or, in some embodiments, between about 2,000 daltons and 5,000 daltons. 
     In some embodiments, the presently disclosed transfection complexes include one or more biologically active agents to be delivered to a cell or to a target tissue in vitro or in vivo. Suitable biologically active agents include, but are not limited to, any molecule that is capable of forming a transfection complex with the presently described transfection reagents and that elicits a biological response when delivered to the interior of a cell or cells or to a tissue in vivo or in vitro. In some embodiments, biologically active agents contemplated for use in the presently described embodiments are cationic, neutral or anionic agents. In some embodiments, the biologically active agents suitable for formulation in the presently disclosed transfection complexes include a protein, DNA or RNA molecule, either alone or in combination with other protein, DNA or RNA molecules in various combinations, though are not limited to; nucleic acids (including but not limited to single or double stranded linear or circular DNA molecules including cDNA molecules, single or double stranded RNA molecules, mRNA, modified mRNA that has increase stability, small interfereing RNA (siRNA) molecules, small hairpin RNA (shRNA) molecules, guideRNA (gRNA), Cas9 protein, Cas9 protein/guide RNA, Cas9DNA/guideRNA, Cas9mRNA/guideRNA, Cas9mRNA/gRNA/ssDNA/RecAprotein, Cas9mRNA/gRNA/ssDNA/recombination protein, Cas9 protein/gRNA/ssDNA/RecAprotein, Cas9 protein/gRNA/ssDNA/recombination protein, microRNA (miRNA) molecules, oligonucleotides, anti-sense oligonucleotides, sense oligonucleotides), polypeptides, antibodies, oligopeptides, therapeutic peptides or protein molecules, peptide nucleic acids (PNAs), cationic, anionic or neutral organic molecules or drugs, in addition to pharmaceutically acceptable salts thereof. In another embodiment, nutrients required for cell growth or nutrients that can be used to enhance protein expression can be delivered into cells by transfection complexes disclosed herein. 
     In certain non-limiting illustrative embodiments, the transfection complexes disclosed herein deliver nucleic acid molecules into cells or tissues in vitro or in vivo, including the delivery of RNA interference molecules (RNAi) or small interfering RNA molecules (siRNA, shRNA or miRNA) into cells for inhibition of gene expression. 
     In some embodiments, the cell surface ligands, the plant virus movement proteins, peptide fragments thereof, or the presently disclosed transfection complexes are used to deliver mRNA molecules or mixtures of mRNA and DNA molecules into a cell or a tissue in vivo or in vitro to promote the expression of a specific protein or proteins. mRNA reprogramming molecules or telomerase are non-limiting examples of mRNA molecules. Cas9 mRNA, and DNA molecules that code for gRNA, CRE mRNA and LoxP containing DNA molecules, SV40T antigen mRNA and DNA molecules with the SV40 origin of replication are non-limiting examples of mRNA and DNA pairs that have utility. Preformed transfection complexes that contain mRNA are non-limiting illustrative embodiments of transfection complexes disclose herein. Telomerase mRNA transfection complexes or Telomerase mRNA and SV40 Large T-antigen complexes as a media supplement are disclosed herein. In some embodiments, preformed transfection complexes containing mRNA are made and the DNA molecule is added to the complex at a later time. 
     In some embodiments, the cell surface ligands, plant virus movement proteins or peptide fragments thereof, or the presently disclosed transfection complexes are used to deliver DNA molecules (including cDNA molecules) into a cell or a tissue in vivo or in vitro to promote the expression of a specific protein or proteins or to synthesize specific RNA molecules, including but not limited to mRNA molecules or RNAi or miRNA or shRNA or sgRNA molecules are also provided. 
     In some embodiments, the cell surface ligands, plant virus movement proteins, or peptide fragments thereof presently disclosed, are used to deliver proteins or protein nucleic acid complexes into a cell or a tissue in vivo or in vitro to effect the function of the protein, as for example in gene editing. In some embodiments, the transfection complexes described herein contain, one or more surface ligands, one or more fusogenic peptides, one or more nuclear targeting peptide, one or more cationic lipid, or one or more neutral lipid. Non-limiting examples of proteins and protein nucleic acid complexes include, recombinases, CRISPR enzymes, Cre recombinase and Cre fusion proteins, transcription activator like effector nucleases (TALEN), genome editing proteins and CRISPR-Cas9 nuclease/guide RNA complex. 
     In some embodiments, proteins such as RNA polymerase, RNA binding proteins or peptides, and transcription factors are bound to nucleic acids and are delivered to cells with the transfection reagents disclosed herein. In certain embodiments, proteins are made anionic by the addition anionic peptides or anionic polymers designed to attach to the protein or an anionic amino acid is added to the C-terminus or N-terminus of the protein. 
     In some embodiments, the transfection complexes described herein may optionally include one or more fusogenic or cell-penetrating peptides. A fusogenic or cell-penetrating peptide is any peptide molecule that is capable of promoting the fusion of a lipid-containing complex to a cell membrane (either a plasma membrane or an endosomal membrane). A variety of fusogenic or cell-penetrating peptides are known in the art and it is well within the skill level of a practitioner to identify suitable fusogenic or cell-penetrating peptides and condition for the use thereof in the present invention without undue experimentation. 
     In some embodiments, the transfection complexes described herein optionally include one or more transfection helpers or targeting moieties in combination with the cell surface ligands, plant virus movement proteins, or peptide fragment thereof described herein. In some embodiments, the targeting moiety is a peptide, a modified peptide, an antibody, a modified antibody, a receptor molecule, a modified receptor molecule, a single or a double stranded nucleic acid molecule, a modified single or double stranded nucleic acid molecule, a peptide or nucleic acid aptamer, a modified peptide or nucleic acid aptamer, an organic molecule, a polysaccharide, an exosome, or any other molecule that is capable of targeting a transfection complex to specific tissue or cell type for targeted delivery of a biologically agent thereto, such as will be readily apparent to those having ordinary skill level in the art. In some embodiments, modification of a peptide, an antibody, a nucleic acid, an aptamer, and the like includes conjugating the peptide, antibody, nucleic acid, aptamer, and the like to a PEG moiety. Alternatively, said modification includes conjugating the peptide, antibody, nucleic acid, aptamer, and the like to a PEG-lipid moiety. A variety of targeting moieties are widely known to those skilled in the art, and all are contemplated for use with the presently described embodiments, without limitation. 
     In some embodiments, the transfection complexes disclosed herein are stable for up to one year and are either contacted with the cells or tissues to be transfected, or are administered to a subject immediately or shortly after being formed. In some embodiments, the transfection complexes disclosed herein are optionally stored for a period of time prior to being contacted with the cells or tissues, or being administered to a subject. The transfection complexes are stable and may be stored for a time period of at least 30 minutes, at least 45 minutes, at least 1 hour, at least 2 hours, at least 3 hours, at least 4 hours, at least 5 hours, at least 10 hours, at least 15 hours, at least 20 hours, at least 24 hours, at least 48 hours, at least 72 hours, at least 5 days, at least 7 days, at least 14 days, at least 28 days, at least 1 month, at least 2 months, at least 3 months, at least 4 months, at least 5 months, at least 6 months or at least 1 year at room temperature, or at a temperature greater than freezing, up to about room temperature. In some embodiments, the formulations described herein include one or more stabilizing agents, preservatives, buffers, and the like, that aid in the long-term stabilization and storage of bioactive formulation, such as will be readily understood by the skilled practitioner of the biological and pharmaceutical arts, and without requiring undue experimentation to achieve. It is also understood, that the storage period can be between any of the aforesaid time periods, for example between 31 minutes and 1 hour or between 1 hour and 24 hours. 
     In another aspect, disclosed herein are methods for the preparation of functional transfection complexes containing cell surface ligands, plant virus movement proteins, or peptides derived thereform, the complexes being as described herein. In some embodiment the functional transfection complex may be used for the preparation of synthetic liposomes or exosomes that contain biologically active macromolecules. In some embodiments, the methods include the step of forming a liposomes or exosome composition containing one or more plant virus movement proteins or peptide fragments or from surface ligands described herein, optionally in combination with one or more helper lipids, stabilizing lipids, transfection helpers, exosomal lipids or lipid extracts, pegylated lipids, targeting moieties, fusion agents, cell penetration agent, lysosomotropic agent. In some embodiments, the methods include the step of forming a lipid-aggregate by encapsulating a biologically active agent in a composition containing one or more surface ligands, plant virus proteins, or peptide fragments thereof described herein, optionally in combination with one or more helper lipids, stabilizing lipids, transfection helpers, pegylated lipids, targeting moieties, fusion agents, lysosomotropic agent and/or exosomes. In some embodiments, the methods alternatively include: 1) mixing one or more surface ligands, plant virus movement proteins, or peptide fragments thereof, with one or more transfection compounds, which optionally include one or more helper lipids, exosomes, surface ligands, stabilizing lipids, transfection helpers, targeting moieties, fusion agents, lysosomaotropic agent, optionally with one or more pegylated lipids, or a salt thereof, in an, an aqueous, alcohol/aqueous, or alcohol solution wherein the alcohol concentration is &lt;10%, &lt;25%, &lt;50%, or &lt;99%; 2) mixing one or more surface ligands, plant virus movement proteins, or peptide fragments thereof, with one or more transfection compounds, which optionally include one or more helper lipids, exosomes, stabilizing lipids, transfection helpers, targeting moieties, surface ligands, fusion agents, cell penetration agents, lysosomotropic agent, and one or more pegylated lipids, or a salt thereof, in a molar percentage such that the one or more transfection compounds are present at 1%-90%; 3) mixing one or more surface ligands, plant virus movement protein, or peptide fragment thereof, with one or more transfection compounds, which optionally include one or more helper lipid, exosomes, stabilizing lipids, transfection helpers, targeting moieties, surface ligands, fusion agents, lysosomotropic agent, one or more pegylated lipids, or a salt thereof, in a molar percentage such that the Pegylated lipids are present at &lt;50%; and 4) mixing one or more surface ligands, plant virus movement proteins, or peptides derived therefrom, with one or more transfection compounds, which optionally include one or more helper lipid, exosomes, stabilizing lipids, transfection helpers, targeting moieties, surface ligands fusion agents, cell penetration agents, lysosomotropic agent, one or more pegylated lipids, or a salt thereof, wherein the pegylated lipid has a polyethylene glycol molecular weight of about 2000-12000 and a fatty acid chain length of C6-C20 alkyl, or C 10 -C 20  alkenyl; and complexing the lipid aggregate in an aqueous, alcohol/aqueous, or alcohol solution with the bioactive agent to form a transfection complex, wherein the alcohol concentration is &lt;50%, preferably less than 40% if pegylated lipids are present. In some embodiments, the alcohol is ethanol. In some embodiments, the alcohol is a pharmaceutically acceptable alcohol such as an alcohol that is liquid at about room temperature, for example, ethanol, propylene glycol, 2-(2-ethoxyethoxy)ethanol (Transcutol™), benzyl alcohol, glycerol, polyethylene glycol 200, polyethylene glycol 300, polyethylene glycol 400 or a mixture thereof. In some embodiments, the alcohol for mixing is different than the alcohol for complexing. Formulations of lipid aggregates as provided above can be provided in 0% to 100% ethanol. In some embodiments, the helper lipid is a neutral lipid. The ratio of cationic to neutral lipid can vary from 100% to 0.1% cationic lipid. 
     In another embodiment preformed lipid aggregates, exosomes, cationic exosomes or liposomes containing cationic, anionic, and or neutral lipids are mixed with cell surface ligands, plant virus movement proteins, or peptides derived therefrom describe herein. In certain embodiments, the lipid aggregates, exosomes or liposomes are optionally formulated with one or more transfection enhancers such as helper lipids, stabilizing lipids, transfection helpers, pegylated lipids, other targeting moieties, cell surface ligands, fusion agents, cell penetration agents, and or lysosomotropic agents. In some embodiments, the cell surface ligands, plant virus movement proteins, or peptides derived therefrom as described herein are added to these formulations at any point prior to or after the bioactive agent is loaded into the transfection complex. 
     In another embodiment a mixture of cationic lipid, neutral lipids and or total lipid extract of exosomes are dissolved in organic solvent such as chloroform and mixed with aqueous solutions optionally containing buffers and one or more cell surface ligands, plant virus movement proteins, or peptides derived therefrom, and optionally one or more transfection enhancers, one or more fusion agents, one or more cell penetration agents, one or more nuclear localization agents and subjected to reveres evaporation to remove the organic solvent leaving behind a lipid aggregate or liposome in solution. 
     In another embodiment a mixture of cationic lipid, neutral lipids and or total lipid extract of exosomes are dissolved in organic solvent such as chloroform or ethanol and mixed with aqueous solutions optionally containing buffers and one or more plant virus movement proteins or peptides derived from plant virus movement proteins, and optionally one or more transfection enhancers, one or more fusion agents, one or more nuclear localization agents, one or more cell surface ligand and subjected to reveres evaporation to remove the organic solvent leaving behind a lipid aggregate or liposome in solution. 
     In another embodiment, a mixture of cationic lipids and, lipids (total lipid extract of exosomes, lipids from exosomes, neutral lipids) in a water, alcohol or in a alcohol water mixture is added to an alcohol solution or alcohol water solution containing buffers and plant virus movement proteins or peptides derived from plant virus movement proteins, and optionally one or more transfection enhancers, one or more exosomes, one or more fusion agents, one or more nuclear localization agents, one or more surface ligands and this solution is optionally micro fluidization, extruded or sonicated to form lipid aggregates or liposomes. 
     In another embodiment, a mixture of cationic lipids and neutral lipid in a water, alcohol or in an alcohol/water mixture is added to an aqueous solution containing buffers and one or more cell surface ligands, plant virus movement proteins, or peptides derived therefrom, and optionally one or more transfection enhancers, one or more exosomes, one or more fusion agents, one or more cell penetration agents, one or more nuclear localization agents one or more surface ligands and this solution is optionally micro fluidization, extruded or sonicated to form lipid aggregates or liposomes. 
     In another aspect, disclosed herein are methods for screening for a tissue-based delivery of a transfection complex or cell type. In some embodiments, the method comprises the step of preparing a plurality of transfection complexes, each transfection complex having at least one cell surface ligand, plant virus movement protein, or peptides derived therefrom, in combination with at least one nucleic acid that facilitates detection of delivery to a tissue or cell type. In some embodiments, the nucleic acid is an RNA molecule or a DNA molecule that encodes a protein that can be directly detected (such as, e.g., Green Fluorescent Protein (GFP), Red Fluorescent Protein, Luciferase, or the like), or encode a protein that effects expression of a protein that can be directly detected. 
     In an embodiment, a method for screening for a cell based or tissue-based delivery of at least one cell surface ligand, plant virus movement protein, or peptide derived therefrom, comprises the step of preparing a plurality of unique transfection complexes, each transfection complex having at least cell surface ligand, plant virus movement protein, or peptide derived therefrom in combination with an mRNA or a cDNA that encodes the detectable protein or a specific transcription factor. Each unique transfection complex is delivered either to cells in culture, intravenously, subcutaneously, or to a tissue to a subject. After a predetermined amount of time, cells in culture or tissues from the subject are harvested and the expression of the detectable protein in various tissues is detected by gross examination, histological examination or by molecular detection (PCR, Western blotting, or the like), or imaged in vivo using the IVIS® Imaging System (Caliper), to determine to which tissue or tissues the transfection complexes containing specific transfection compounds are delivered. 
     In an embodiment, a method for screening cells in culture or tissue-based delivery of a transfection complex comprises the step of preparing a plurality of unique transfection complexes, each transfection complex having at least one test cell surface ligand, plant virus movement protein, or peptide derived therefrom in combination with an mRNA or a cDNA that encodes a specific transcription factor. Each unique transfection complex may be delivered to cells in culture, intravenously, subcutaneously, or to a tissue to a transgenic animal that expresses a reporter gene (such as, e.g., luciferase) under the control of the specific transcription factor. After a predetermined amount of time, tissues from the transgenic animal may be harvested and the expression of reporter gene in various tissues may be detected by gross examination, histological examination or by molecular detection (PCR, Western blotting, or the like). If the reporter gene is luciferase, detection may be accomplished in-vivo using the IVIS® Imaging System (Caliper). 
     In some embodiments, cell surface ligand, plant virus movement protein, or peptide derived therefrom of the presently disclosed transfection complexes are used to deliver exosomes into a cell or a tissue in vivo or in vitro to effect the function of the biological cargo in the exosomes. In some embodiments, the transfection complex described herein comprise one or more fusogenic peptides, one or more cell penetration agents, one or more nuclear targeting peptide, one or more cationic lipid, one or more plant virus movement proteins or peptides derived from plant virus movement proteins, one or more surface ligands or one or more neutral lipids. In certain embodiments, the transfection complexes described herein comprise one or more of the cationic lipids described in Formula I. 
     In another aspect, disclosed herein are compositions and methods that provide improved efficiency for introducing molecules and macromolecules, such as nucleic acids, proteins, peptides, nutrients and pharmaceuticals into cells. Accordingly, provided herein are compositions comprising a nucleic acid molecule, a transfection agent and a transfection enhancer. 
     In some embodiments, the transfection enhancer is a surface ligand that comprises amino acid sequences derived from a cell binding adhesion proteins. Collagen, fibronectin, lamin, veronectin, cadherin, nidogen, fibrinogen, elastin, bone asialoprotein, osteopontin and tenascin-C are non-limiting examples of cell binding adhesion proteins. In some embodiments, the surface ligand is the above-listed full-length protein. In other embodiments, the surface ligand is a fragment of the above-listed protein having greater than 5 amino acids in length. In other embodiments, the length of the fragment of the above-listed protein is greater than 5 amino acids is greater than 10 amino acids, greater than 15 amino acids, greater than 20 amino acids, greater than 25 amino acids, greater than 30 amino acids, greater than 35 amino acids, or greater than 40 amino acids. 
     In some embodiments, cell surface ligand, plant virus movement protein, or peptide derived therefrom proteins and describe herein comprise a nucleic acid binding moiety functionally linked to the amino acid sequence of the cell surface ligand, plant virus movement protein, or peptide derived therefrom proteins. Suitable nucleic acid binding moieties include, but are not limited to, a polycationic peptide sequence, a polyamine, a peptide nucleic acid, spermine, spermidine, carboxyspermidine, carboxy spermine, spermine and spermidine analogs, nucleic acid intercalaters, and the like. In certain embodiments, the nucleic acid binding moiety is covalently linked to the transfection promoting cell surface ligand comprising adhesion protein amino acid sequences. In further embodiments, the transfection agent is a cationic lipid, such as those described below, a polyamine, a polycationic peptide sequence, a cationic dendrimer, or the like. In some embodiments, the cell surface ligand adhesion sequence or peptide derived from a plant movement protein is a multimer of itself or other adhesion sequences. In certain embodiments, the cell binding adhesion or peptide derived from a plant movement protein amino sequence is cyclized. In other embodiments, the surface ligands or peptide derived from a plant movement protein also contain other peptide sequence that enhance transfection efficiency, such as linkers, spacers, or nuclear targeting sequences. 
     In some embodiments, cell surface ligand, plant virus movement protein, or peptide derived therefrom described herein are attached directly to the binding molecule by covalent bonding, or are connected to the binding molecule via a spacer. The term “spacer,” or “linker,” which are used interchangeably herein, as used herein refers to a chemical structure that links two molecules to each other. In some embodiments, the spacer binds each molecule on a different part of the spacer molecule. In other embodiments, the spacer is a hydrophilic moiety and comprises about 6 to 30 carbon atoms. In other embodiments, the spacer comprises a ployether, for example —CH 2 —O—(CH 2 —CH 2 —O—) i CH 2 -. In other embodiments, the spacer comprises a hydrophilic polymer, for example [(gly) i (ser) j ] k  (SEQ ID NO: 585). In these formulae i ranges from 1 to 6, j ranges from 1 to 6, and k ranges from 3 to 20. In some embodiments, the spacer is a peptide of sequence APYKAWK (SEQ ID NO:505). In other embodiments, the spacer is a sequence that is degraded in vivo by a peptidase. 
     In some embodiments, the cell surface ligand, plant virus movement protein, or peptide derived therefrom described herein are functionally linked to a lipid, such as a cationic or neutral lipid. In some of these embodiments, the linked moiety is used for delivery of macromolecules into cells. For example, a cell surface ligand, plant virus movement protein, or peptide derived therefrom, or a cell binding adhesion peptide sequences amino acid sequence is covalently linked to a lipid, such as a cationic lipid, a lysolipid, using methods known in the art. 
     In certain embodiments, the cell surface ligand, plant virus movement protein, or peptide derived therefrom sequences described herein also are functionally linked to an amino acid sequence that inserts itself into lipid membranes, such as membrane anchor peptides or proteins. In other embodiments, the cell surface ligand peptide sequences are linked to chemical compositions that associate with lipids. 
     In other embodiments, the transfection complexes or liposomal compositions with the cell surface ligand, plant virus movement protein, or peptide derived therefrom described herein also comprise other transfection enhancing agents, such as a nuclear localization protein or peptide, a fusogenic peptide or protein, a transport peptide or protein, a viral peptide or protein, or a lysomoltropic agent. In certain embodiments, the viral peptide is derived from a virus enveloped or non-enveloped virus, for example an influenza virus, a vesicular stomatitis virus, an adenovirus, an alphavirus, a Semliki Forest Virus, a hepatitis virus, a herpes virus, an HIV virus, or a simian virus. In some embodiments, the transfection enhancing agent is, for example, insulin, a transferrin, a epidermal growth factor, a fibroblast growth factor, a cell targeting antibody or fragment from an antibody, a lactoferrin, a fibronectin, an adenovirus penton base, Knob, a hexon protein, a vesicular stomatitis virus glycoprotein, a Semliki Forest Virus core protein, an influenza hemagglutinin, a hepatitis B core protein, an HIV Tat protein, a herpes simplex virus VP22 protein, a histone protein, an arginine rich cell permeability protein, a high mobility group protein, and invasin protein, an internalin protein, an endotoxin, a diptheria toxin, a shigella toxin, a melittin, a magainin, a gramicidin, a cecrophin, a defensin, a protegrin, a tachyplesin, a thionin, a indolicidin, a bactenecin, a drosomycin, an apidaecin, a cathelicidin, a bacteriacidal-permability-increasing protein, a nisin, a buforin, or fragments thereof. In other embodiments, the transfection enhancing agent is chloroquine, a lysosomotrophic compound or combinations thereof. In other embodiments exosomes or exosomal derived lipids or proteins are the transfection enhanceagent. In other embodiments, the transfection enhancer agent comprises multimers of the same or different peptides or proteins. 
     Suitable nuclear localization peptides or proteins included in transfection complexes or liposomal compositions include, but are not limited to, a sequence selected from the group consisting of SEQ ID NOs: 1-41, as set forth in Table 1, below, or in the sequence listings. 
     Proteins such as histones, protamines, HMG proteins, and viral core proteins or coat proteins comprise nuclear localization proteins. In some embodiments, these proteins or fragments thereof are used to enhance transfection. In some embodiments, the nuclear localization peptide is optionally linked to a nucleic acid binding moiety, for example via a covalent linkage. Spacer sequences are optionally used between the DNA binding sequence and the nuclear localization sequence. In some embodiments, the nuclear localization sequences are linked to helper lipids or other peptides proteins or compounds that associate with lipid bilayers. 
     In some embodiments, the compositions described herein also comprise a fusion agent or combinations of fusion agents, which in some embodiments also function as an amphipathic peptide. Suitable fusion peptides include, but are not limited to, a sequence selected from the group consisting of SEQ ID NOs:42-92, as set forth in Table 1, below, or in the sequence listings. 
     In some embodiments, the fusion agent is optionally linked to a nucleic acid binding moiety, for example via a covalent linkage. The peptides KK, KKK, KKKK (SEQ ID NO:97), RR, RRR, RRRR (SEQ ID NO:105) can be linked to fusion agents of SEQ ID NOs:42-92. In certain embodiments, fusion peptides are linked to helper lipids, cationic lipids, or other peptides or proteins that associate with lipid bilayers. Spacer sequences are optionally used between the DNA binding sequence and the fusion agent sequence 
     In certain embodiments, the compositions disclosed herein comprise a cell penetration agent or combinations of cell penetration agents. Suitable cell penetration agents include, but are not limited to, a sequence selected from the group consisting of SEQ ID NOs:93-96 as set forth in Table 1, below, or in the sequence listings. 
     In some embodiments, the cell penetration agents are optionally linked to a nucleic acid binding moiety, for example via a covalent linkage. In other embodiments, the cell penetration agents are linked to helper lipids or other peptides or proteins that associate with lipid bilayers. 
     In some embodiments, the nuclear localization sequences, the fusion agents, cell surface ligand or the cell penetration agents are linked to the GPI anchor peptides, the sequence FTLTGLLGTLVTMGLLT (SEQ ID NO:504) being a non limiting example. 
     In some embodiments, the nucleic acid binding moieties that are linked to different transfection enhancer and are part of transfection complexes have different binding affinity for nucleic acids depending on the needed functionality for attachment, condensation of nucleic acid, and the rate of release of nucleic acid from the nucleic acid binding moiety. Suitable nucleic acid binding moieties include, but are not limited to a polycationic peptide sequence, a polyamine, a peptide nucleic acid, spermine, spermidine, carboxyspermidine, carboxy spermine, spermine and spermidine analogs, nucleic acid intercalaters, and the like. 
     In some embodiments, the compositions described herein comprise combinations of different transfection enhancers with different nucleic acid binding moieties. Suitable nucleic acid binding peptides include, but are not limited to a sequence of SEQ ID NOs:97-149, as set forth in Table 1, below, or in the sequence listings. 
     In some embodiments, the nucleic acid binding moieties also serve as transfection enhancers when bound to nucleic acids, or alternatively serve as condensing agents. Suitable nucleic acid condensing peptides include, but are not limited to, a sequence selected from the group consisting of the peptides of SEQ ID NOs:97-149 as set forth in Table 1, below, or in the sequence listings. In some embodiments, multimers of these peptides are also synthesized and used as condensing agents. In some embodiments, nuclear localization sequences are also used as condensing agents if they have enough cationic charge. 
     Suitable nucleic acid binding moieties include, but are not limited to a polycationic peptide sequence, a polyamine, a peptide nucleic acid, spermine, spermidine, carboxyspermidine, carboxy spermine, spermine and spermidine analogs, nucleic acid intercalaters, and the like 
     One skilled in the art will readily recognize that the surface ligand chosen depends on which receptor is being bound. Since different types of cells have different receptors, this provides a method of targeting nucleic acid, peptides, protein, and compounds to specific cell types, depending on which cell surface ligand is used. Thus, the preferred cell surface ligand or ligands may depend on the targeted cell type. 
     In some embodiments, the transfection enhancers that are used in combination with the cell surface ligand, plant virus movement protein, or peptide derived therefrom disclose herein include, but are not limited to, the peptides or proteins selected from the group consisting of a collagen, a fibronectin, a lamin, a veronectin, a cadherin, a nidogen, a fibrinogen, a elastin, a bone asialoprotein, a osteopontin, a tenascin-C, Avadin, insulin, a transferrin, a epidermal growth factor, a fibroblast growth factor, a cell targeting antibody, a lactoferrin, an enveloped virus, a non-enveloped virus, an adenovirus penton base, a knob protein, a hexon protein, a vesicular stomatitis virus glycoprotein, a Semliki Forest Virus core protein, an influenza hemagglutinin, a hepatitis B core protein, an HIV Tat protein, a herpes simplex virus VP22 protein, a histone protein, an arginine rich cell permeability protein, a high mobility group protein, invasin protein, internalin protein, an endotoxin, a non-toxic diptheria toxin, a non-toxic shigella toxin, a melittin, a magainin, a gramicidin, a cecrophin, a defensin, a protegrin, a tachyplesin, a thionin, a indolicidin, a bactenecin, a drosomycin, an apidaecin, a cathelicidin, a bacteriacidal-permability-increasing protein, a nisin, a buforin, a fragment thereof, and a sequence selected from the group consisting of SEQ ID NOs: 150-503, as set forth in Table 1, below, or in the sequence listings. 
     In some embodiments, the transfection enhancing agent is chloroquine, a lysosomotrophic compound or combinations thereof. In certain embodiments, the transfection enhancer agent comprises multimers of the same or different peptide enhancers, protein or protein fragments of transfection enhancers. 
     In some embodiments, the aforementioned peptides are optionally linked to a moiety selected from the group consisting of a nucleic acid binding moiety, a helper lipid, a cationic lipid, a cationic polymer, and a GPI anchor peptide. 
     In certain embodiments, the aforementioned peptides are optionally linked to a chemical moiety of Formula I, 
     
       
         
         
             
             
         
       
     
     or a pharmaceutically acceptable salt thereof, where
         W 1 ═W 2 ═CH 2 —N(R 3 R 4 ), R 1 ═R 2 ═C 8 -C 22  alkyl, X═O; R 3 ═CH 3 , R 4 ═—C(═O)—C(NH 2 )—CH 2 —(CH 2 )n-NH 2 , n=0-6, y=0; or   W 1 ═W 2 ═CH 2 —N(R 3 R 4 ), R 1 ═R 2 ═C 8 -C 22  alkyl, X═O; R 3 ═CH 3 , R 4 ═—C(═O)—C(NH 2 )—CH 2 —(CH 2 )n-OH, n=0-6, y=0; or   W 1 ═W 2 ═CH 2 —N(R 3 R 4 ), R 1 ═R 2 ═C 8 -C 22  alkyl, X═O; R 3 ═CH 3 , R 4 ═—C(═O)—C(OH)—CH 2 —(CH 2 )n-NH 2 , n=0-6, y=0; or   W 1 ═W 2 ═CH 2 —N(R 3 R 4 ), R 1 ═R 2 ═C 8 -C 22  alkyl, X═O; R 3 ═CH 3 , R 4 ═—C(═O)—C(NH-iso-butyl)-CH 2 —(CH 2 )n-O-iso-butyl, n=0-6, y=0; or   W 1 ═W 2 ═CH 2 —N(R 3 R 4 ), R 1 ═R 2 ═C 8 -C 22  alkyl, X═O; R 3 ═CH 3 , R 4 ═—C(═O)—C(NH 2 )—CH 2 —(CH 2 )n-C(═NH)—NH 2 , n=0-6, y=0; or   W 1 ═W 2 ═CH 2 —N(R 3 R 4 ), R 1 ═R 2 ═C 8 -C 22  alkyl, X═O; R 3 ═CH 3 , R 4 ═—C(═O)—C(NH 2 )—CH 2 —(CH 2 )n-His, n=0-6, y=0; or   W 1 ═W 2 ═CH 2 —N(R 3 R 4 ), R 1 ═R 2 ═C 8 -C 22  alkyl, X═O; R 3 ═CH 3 , R 4 ═—C(═O)-spermine, y=0; or   W 1 ═W 2 ═CH 2 —N(R 3 R 4 ), R 1 ═R 2 ═C 8 -C 22  alkyl, X═O; R 3 ═CH 3 ; R 4 ═H, y=0; or   W 1 ═W 2 ═CH 2 —N(R 3 R 4 ), R 1 ═R 2 ═C 8 -C 22  alkyl, X═O; R 3 ═CH 3 ; R 4 ═CH 2 (CH 2 ) m —NH 2 ; m=1-6, y=0; or   W 1 ═W 2 ═CH 2 —N(R 3 R 4 ), R 1 ═R 2 ═C 8 -C 22  alkyl, X═O; R 3 ═CH 3 ; R 4 ═CH 2 (CH 2 ) m —NH—C(═O)-spermine; m=1-6, y=0; or   W 1 ═W 2 ═CH 2 —N(R 3 R 4 ), R 1 ═R 2 ═C 8 -C 22  alkyl, X═O; R 3 ═CH 3 ; R 4 ═CH 2 (CH 2 ) m —NH—(C═O)-amino acid side chain; m=1-6, y=0; or   W 1 ═W 2 ═CH 2 —N(R 3 R 4 ), R 1 ═R 2 ═C 8 -C 22  alkyl, X═O; R 3 ═CH 3 ; R 4 ═(CH 2 ) m (CH—OH)(CH 2 ) m —NH 2 , y=0; or   W 1 ═W 2 ═CH 2 —N(R 3 R 4 ), R 1 ═R 2 ═C 8 -C 22  alkyl, X═O; R 3 ═CH 3 ; R 4 ═CH 2 (CH 2 )n-NH 2 , y=0; or   W 1 ═H, W 2 ═CH 2 —N(R 3 R 4 )CH 2 —R 5 ; R 3 ═R 4 ═CH 3 ; R 5 ═H, y=0; or   W 1 ═H; W 2 ═CH 2 —O—P(═O)(OMe)-O—CH 2 CH 2 —NH—C(═O)-spermine, y=0; or   W 1 ═H; W 2 ═CH 2 —O—P(═O)(OMe)-O—CH 2 CH 2 —NH—C(═O)-amino acid side chain, y=0; or   W 1 ═H; W 2 ═CH 2 —O—P(═O)(OMe)-O—CH 2 CH 2 —N + (CH 3 ) 3 Cl, y=0; or   W 1 ═H; W 2 ═CH 2 —O—P(═O)(O)—O—CH 2 CH 2 —NH—C(═O)-spermine, y=0; or   Z═(CH 2 ) q , W 1 ═W 2 ═CH 2 —N(R 3 R 4 ), R 1 ═R 2 ═C 8 -C 22  alkyl, X═O; R 3 ═CH 3 , R 4 ═—C(═O)—C(NH 2 )—CH 2 —(CH 2 )n-NH 2 , n=0-6,q=1-3, y=1; or   Z═(CH 2 ) q , W 1 ═W 2 ═CH 2 —N(R 3 R 4 ), R 1 ═R 2 ═C 8 -C 22  alkyl, X═O; R 3 ═CH 3 , R 4 ═—C(═O)—C(NH 2 )—CH 2 —(CH 2 ) n —OH, n=0-6,q=1-3, y=1; or   Z═(CH 2 ) q , W1═W 2 ═CH 2 —N(R 3 R 4 ), R 1 ═R 2 ═C 8 -C 22  alkyl, X═O; R 3 ═CH 3 , R 4 ═—C(═O)—C(OH)—CH 2 —(CH 2 ) n —NH 2 , n=0-6, q=1-3, y=1; or   Z═(CH 2 ) q , W 1 ═W 2 ═CH 2 —N(R 3 R 4 ), R 1 ═R 2 ═C 8 -C 22  alkyl, X═O; R 3 ═CH 3 , R 4 ═—C(═O)—C(NH-iso-butyl)-CH 2 —(CH 2 ) n —O-iso-butyl, n=0-6,y=1; or   Z═(CH 2 ) q , W 1 ═W 2 ═CH 2 —N(R 3 R 4 ), R 1 ═R 2 ═C 8 -C 22  alkyl, X═O; R 3 ═CH 3 , R 4 ═—C(═O)—C(NH 2 )—CH 2 —(CH 2 ) n —C(═NH)—NH 2 , n=0-6, q=1-3, y=1; or   Z═(CH 2 ) q , W 1 ═W 2 ═CH 2 —N(R 3 R 4 ), R 1 ═R 2 ═C 8 -C 22  alkyl, X═O; R 3 ═CH 3 , R 4 ═—C(═O)—C(NH 2 )—CH 2 —(CH 2 ) n -His, n=0-6, q=1-3, y=1; or   Z═(CH 2 ) q W 1 ═W 2 ═CH 2 —N(R 3 R 4 ), R 1 ═R 2 ═C 8 -C 22  alkyl, X═O; R 3 ═CH 3 , R 4 ═—C(═O)-spermine, q=1-3, y=1; or   Z═(CH 2 ) q , W 1 ═W 2 ═CH 2 —N(R 3 R 4 ), R 1 ═R 2 ═C 8 -C 22  alkyl, X═O; R 3 ═CH 3 ; R 4 ═H, q=1-3, y=1; or   Z═(CH 2 ) q W 1 ═W 2 ═CH 2 —N(R 3 R 4 ), R 1 ═R 2 ═C 8 -C 22  alkyl, X═O; R 3 ═CH 3 ; R 4 ═CH 2 (CH 2 ) m —NH 2 ; m=1-6, q=1-3, y=1; or   Z═(CH 2 ) q , W 1 ═W 2 ═CH 2 —N(R 3 R 4 ), R 1 ═R 2 ═C 8 -C 22  alkyl, X═O; R 3 ═CH 3 ; R 4 ═CH 2 (CH 2 ) m —NH—C(═O)-spermine; m=1-6, q=1-3, y=1; or   Z═(CH 2 ) q , W 1 ═W 2 ═CH 2 —N(R 3 R 4 ), R 1 ═R 2 ═C 8 -C 22  alkyl, X═O; R 3 ═CH 3 ; R 4 ═CH 2 (CH 2 ) m —NH—(C═O)-amino acid side chain; m=1-6, q=1-3, y=1; or   Z═(CH 2 ) q , W 1 ═W 2 ═CH 2 —N(R 3 R 4 ), R 1 ══C 8 -C 22  alkyl, X═O; R 3 ═CH 3 ; R 4 ═(CH 2 ) m (CH—OH)(CH 2 ) m —NH 2 , q=1-3, y=1; or   Z═(CH 2 ) q , W 1 ═W 2 ═CH 2 —N(R 3 R 4 ), R 1 ══C 8 -C 22  alkyl, X═O; R 3 ═CH 3 ; R 4 ═CH 2 (CH 2 ) n —NH 2 , q=1-3, y=1; or   Z═(CH 2 ) q —N(R 3 )—(CH 2 ) q , W 1 ═W 2 ═CH 2 —N(R 3 R 4 ), R 1 ═R 2 ═C 8 -C 22  alkyl, X═O; R 3 ═CH 3 , R 4 ═—C(═O)—C(NH 2 )—CH 2 —(CH 2 )n-NH 2 , n=0-6, q=1-3, y=1; or   Z═(CH 2 ) p —N(R 3 )—(CH 2 ) p , W 1 ═W 2 ═CH 2 —N(R 3 R 4 ), R 1 ═R 2 ═C 8 -C 22  alkyl, X═O; R 3 ═CH 3 , R 4 ═—C(═O)—C(NH 2 )—CH 2 —(CH 2 )n-OH, n=0-6, q=1-3, y=1; or   Z═(CH 2 ) q —N(R 3 )—(CH 2 ) q , W1═W 2 ═CH 2 —N(R 3 R 4 ), R 1 ═R 2 ═C 8 -C 22  alkyl, X═O; R 3 ═CH 3 , R 4 ═—C(═O)—C(OH)—CH 2 —(CH 2 )n-NH 2 , n=0-6, q=1-3, y=1; or   Z═(CH 2 ) q —N(R 3 )—(CH 2 ) q , W 1 ═W 2 ═CH 2 —N(R 3 R 4 ), R 1 ═R 2 ═C 8 -C 22  alkyl, X═O; R 3 ═CH 3 , R 4 ═—C(═O)—C(NH-iso-butyl)-CH 2 —(CH 2 )n-O-iso-butyl, n=0-6,y=1; or   Z═(CH 2 ) q —N(R 3 )—(CH 2 ) q , W 1 ═W 2 ═CH 2 —N(R 3 R 4 ), R 1 ═R 2 ═C 8 -C 22  alkyl, X═O; R 3 ═CH 3 , R 4 ═—C(═O)—C(NH 2 )—CH 2 —(CH 2 )n-C(═NH)—NH 2 , n=0-6, q=1-3, y=1; or   Z═(CH 2 ) q —N(R 3 )—(CH 2 ) q , W 1 ═W 2 ═CH 2 —N(R 3 R 4 ), R 1 ═R 2 ═C 8 -C 22  alkyl, X═O; R 3 ═CH 3 , R 4 ═—C(═O)—C(NH 2 )—CH 2 —(CH 2 )n-His, n=0-6, q=1-3, y=1; or   Z═(CH 2 ) q —N(R 3 )—(CH 2 ) q , W 1 ═W 2 ═CH 2 —N(R 3 R 4 ), R 1 ═R 2 ═C 8 -C 22  alkyl, X═O; R 3 ═CH 3 , R 4 ═—C(═O)-spermine, q=1-3, y=1; or   Z═(CH 2 ) q —N(R 3 )—(CH 2 ) q , W 1 ═W 2 ═CH 2 —N(R 3 R 4 ), R 1 ═R 2 ═C 8 -C 22  alkyl, X═O; R 3 ═CH 3 ; R 4 ═H, q=1-3, y=1; or   Z═(CH 2 ) q —N(R 3 )—(CH 2 ) q , W 1 ═W 2 ═CH 2 —N(R 3 R 4 ), R 1 ═R 2 ═C 8 -C 22  alkyl, X═O; R 3 ═CH 3 ; R 4 ═CH 2 (CH 2 ) m —NH 2 ; m=1-6, q=1-3, y=1; or   Z═(CH 2 ) q —N(R 3 )—(CH 2 ) q , W 1 ═W 2 ═CH 2 —N(R 3 R 4 ), R 1 ═R 2 ═C 8 -C 22  alkyl, X═O; R 3 ═CH 3 ; R 4 ═CH 2 (CH 2 ) m —NH—C(═O)-spermine; m=1-6, q=1-3, y=1; or   Z═(CH 2 ) q —N(R 3 )—(CH 2 ) q , W 1 ═W 2 ═CH 2 —N(R 3 R 4 ), R 1 ═R 2 ═C 8 -C 22  alkyl, X═O; R 3 ═CH 3 ; R 4 ═CH 2 (CH 2 ) m —NH—(C═O)-amino acid side chain; m=1-6, q=1-3, y=1; or   Z═(CH 2 ) q —N(R 3 )—(CH 2 ) q , W 1 ═W 2 ═CH 2 —N(R 3 R 4 ), R 1 ═R 2 ═C 8 -C 22  alkyl, X═O; R 3 ═CH 3 ; R 4 ═(CH 2 ) m (CH—OH)(CH 2 ) m —NH 2 , q=1-3, y=1; or   Z═(CH 2 ) q —N(R 3 )—(CH 2 ) q , W 1 ═W 2 ═CH 2 —N(R 3 R 4 ), R 1 ═R 2 ═C 8 -C 22  alkyl, X═O; R 3 ═CH 3 ; R 4 ═CH 2 (CH 2 ) n —NH 2 , q=1-3, y=1; or   Z═(CH 2 ) q —S—S—(CH 2 ) q , W 1 ═W 2 ═CH 2 —N(R 3 R 4 ), R 1 ═R 2 ═C 8 -C 22  alkyl, X═O; R 3 ═CH 3 , R 4 ═—C(═O)—C(NH 2 )—CH 2 —(CH 2 ) n —NH 2 , n=0-6, q=1-3, y=1; or   Z═(CH 2 ) q —S—S—(CH 2 ) q , W 1 ═W 2 ═CH 2 —N(R 3 R 4 ), R 1 ═R 2 ═C 8 -C 22  alkyl, X═O; R 3 ═CH 3 , R 4 ═—C(═O)—C(NH 2 )—CH 2 —(CH 2 ) n —OH, n=0-6, q=1-3, y=1; or   Z═(CH 2 ) q —S—S—(CH 2 ) q , W1═W 2 ═CH 2 —N(R 3 R 4 ), R 1 ═R 2 ═C 8 -C 22  alkyl, X═O; R 3 ═CH 3 , R 4 ═—C(═O)—C(OH)—CH 2 —(CH 2 ) n —NH 2 , n=0-6, q=1-3, y=1; or   Z═(CH 2 ) q —S—S—(CH 2 ) q , W 1 ═W 2 ═CH 2 —N(R 3 R 4 ), R 1 ═R 2 ═C 8 -C 22  alkyl, X═O; R 3 ═CH 3 , R 4 ═—C(═O)—C(NH-iso-butyl)-CH 2 —(CH 2 ) n —O-iso-butyl, n=0-6,y=1; or   Z═(CH 2 ) q —S—S—(CH 2 ) q , W 1 ═W 2 ═CH 2 —N(R 3 R 4 ), R 1 ═R 2 ═C 8 -C 22  alkyl, X═O; R 3 ═CH 3 , R 4 ═—C(═O)—C(NH 2 )—CH 2 —(CH 2 ) n —C(═NH)—NH 2 , n=0-6, q=1-3, y=1; or   Z═(CH 2 ) q —S—S—(CH 2 ) q , W 1 ═W 2 ═CH 2 —N(R 3 R 4 ), R 1 ═R 2 ═C 8 -C 22  alkyl, X═O; R 3 ═CH 3 , R 4 ═—C(═O)—C(NH 2 )—CH 2 —(CH 2 ) n -His, n=0-6, q=1-3, y=1; or   Z═(CH 2 ) q —S—S—(CH 2 ) q , W 1 ═W 2 ═CH 2 —N(R 3 R 4 ), R 1 ═R 2 ═C 8 -C 22  alkyl, X═O; R 3 ═CH 3 , R 4 ═—C(═O)-spermine, q=1-3, y=1; or   Z═(CH 2 ) q —S—S—(CH 2 ) q , W 1 ═W 2 ═CH 2 —N(R 3 R 4 ), R 1 ═R 2 ═C 8 -C 22  alkyl, X═O; R 3 ═CH 3 ; R 4 ═H, q=1-3, y=1; or   Z═(CH 2 ) q —S—S—(CH 2 ) q , W 1 ═W 2 ═CH 2 —N(R 3 R 4 ), R 1 ═R 2 ═C 8 -C 22  alkyl, X═O; R 3 ═CH 3 ; R 4 ═CH 2 (CH 2 ) m —NH 2 ; m=1-6, q=1-3, y=1; or   Z═(CH 2 ) q —S—S—(CH 2 ) q , W 1 ═W 2 ═CH 2 —N(R 3 R 4 ), R 1 ═R 2 ═C 8 -C 22  alkyl, X═O; R 3 ═CH 3 ; R 4 ═CH 2 (CH 2 ) m —NH—C(═O)-spermine; m=1-6, q=1-3, y=1; or   Z═(CH 2 ) q —S—S—(CH 2 ) q , W 1 ═W 2 ═CH 2 —N(R 3 R 4 ), R 1 ═R 2 ═C 8 -C 22  alkyl, X═O; R 3 ═CH 3 ; R 4 ═CH 2 (CH 2 ) m —NH—(C═O)-amino acid side chain; m=1-6, q=1-3, y=1; or   Z═(CH 2 ) q —S—S—(CH 2 ) q , W 1 ═W 2 ═CH 2 —N(R 3 R 4 ), R 1 ═R 2 ═C 8 -C 22  alkyl, X═O; R 3 ═CH 3 ; R 4 ═(CH 2 ) m (CH—OH)(CH 2 ) m —NH 2 , q=1-3, y=1; or   Z═(CH 2 ) q —S—S—(CH 2 ) q , W 1 ═W 2 ═CH 2 —N(R 3 R 4 ), R 1 ═R 2 ═C 8 -C 22  alkyl, X═O; R 3 ═CH 3 ; R 4 ═CH 2 (CH 2 ) n —NH 2 , q=1-3, y=1; or   Z═(CH 2 ) q W 1 ═W 2 ═CH 2 —N(R 3 R 4 ), R 1 ═R 2 ═C 8 -C 22  alkyl, X═O; R 3 ═CH 3 ; R 4 ═COCH 2 (OCH 2 CH 2 ) m —NH 2 ; m=1-6, q=1-3, y=0 or 1; or   Z═(CH 2 ) q —S—S—(CH 2 ) q , W 1 ═W 2 ═CH 2 —N(R 3 R 4 ), R 1 ═R 2 ═C 8 -C 22  alkyl, X═O; R 3 ═CH 3 ; R 4 ═COCH 2 (OCH 2 CH 2 ) m —NH 2 ; m=1-6, q=1-3, y=1; or   Z═(CH 2 ) q —N(R 3 )—(CH 2 ) q , W 1 ═W 2 ═CH 2 —N(R 3 R 4 ), R 1 ═R 2 ═C 8 -C 22  alkyl, X═O; R 3 ═CH 3 ; R 4  ═COCH 2 (OCH 2 CH 2 ) m —NH 2 ; m=1-6, q=1-3, y=1; or   W 1 ═W 2 ═—CH 2 —B—CH 2 —N(R 3 R 4 ), R 1 ═R 2 ═C 8 -C 22  alkyl, X═O; R 3 ═CH 3 , R 4 ═—C(═O)—C(NH 2 )—CH 2 —(CH 2 )n-NH 2 , n=0-6, y=0 where B is selected from the group —O(CH 2 ) i , —S(CH 2 ) i , —S—S(CH 2 ) i , —SO 2 —(CH 2 ) i , i=1-5; or   W 1 ═W 2 ═—CH 2 —B—CH 2 —N(R 3 R 4 ), R 1 ═R 2 ═C 8 -C 22  alkyl, X═O; R 3 ═CH 3 , R 4 ═—C(═O)—C(NH 2 )—CH 2 —(CH 2 )n-OH, n=0-6, y=0 where B is selected from the group —O(CH 2 ) i , —S(CH 2 ) i , —S—S(CH 2 ) i , —SO 2 —(CH 2 ) i , i=1-5; or   W 1 ═W 2 ═—CH 2 —B—CH 2 —N(R 3 R 4 ), R 1 ═R 2 ═C 8 -C 22  alkyl, X═O; R 3 ═CH 3 , R 4 ═—C(═O)—C(OH)—CH 2 —(CH 2 )n-NH 2 , n=0-6, y=0 where B is selected from the group —O(CH 2 ) i , —S(CH 2 ) i , —S—S(CH 2 ) i , —SO 2 —(CH 2 ) i , i=1-5; or   W 1 ═W 2 ═—CH 2 —B—CH 2 —N(R 3 R 4 ), R 1 ═R 2 ═C 8 -C 22  alkyl, X═O; R 3 ═CH 3 , R 4 ═—C(═O)—C(NH-iso-butyl)-CH 2 —(CH 2 ) n —O-iso-butyl, n=0-6, y=0 where B is selected from the group —O(CH 2 ) i , —S(CH 2 ) i , —S—S(CH 2 ) i , —SO 2 —(CH 2 ) i , i=1-5; or   W 1 ═W 2 ═—CH 2 —B—CH 2 —N(R 3 R 4 ), R 1 ═R 2 ═C 8 -C 22  alkyl, X═O; R 3 ═CH 3 , R 4 ═—C(═O)—C(NH 2 )—CH 2 —(CH 2 ) n —C(═NH)—NH 2 , n=0-6, y=0 where B is selected from the group —O(CH 2 ) i , —S(CH 2 ) i , —S—S(CH 2 ) i , —SO 2 —(CH 2 ) i , i=1-5; or   W 1 ═W 2 ═—CH 2 —B—CH 2 —N(R 3 R 4 ), R 1 ═R 2 ═C 8 -C 22  alkyl, X═O; R 3 ═CH 3 , R 4 ═—C(═O)—C(NH 2 )—CH 2 —(CH 2 )n-His, n=0-6, y=0 where B is selected from the group —O(CH 2 ) i , —S(CH 2 ) i , —S—S(CH 2 ) i , —SO 2 —(CH 2 ) i , i=1-5; or   W 1 ═W 2 ═—CH 2 —B—CH 2 —N(R 3 R 4 ), R 1 ═R 2 ═C 8 -C 22  alkyl, X═O; R 3 ═CH 3 , R 4 ═—C(═O)-spermine, y=0 where B is selected from the group —O(CH 2 ) i , —S(CH 2 ) i , —S—S(CH 2 ) i , —SO 2 —(CH 2 ) i , i=1-5; or   W 1 ═W 2 ═—CH 2 —B—CH 2 —N(R 3 R 4 ), R 1 ═R 2 ═C 8 -C 22  alkyl, X═O; R 3 ═CH 3 ; R 4 ═H, y=0 where B is selected from the group —O(CH 2 ) i , —S(CH 2 ) i , —S—S(CH 2 ) i , —SO 2 —(CH 2 ) i , i=1-5; or   W 1 ═W 2 ═CH 2 —N(R 3 R 4 ), R 1 ═R 2 ═C 8 -C 22  alkyl, X═O; R 3 ═CH 3 ; R 4 ═CH 2 (CH 2 ) m —NH 2 ; m=1-6, y=0; or   W 1 ═W 2 ═—CH 2 —B—CH 2 —N(R 3 R 4 ), R 1 ═R 2 ═C 8 -C 22  alkyl, X═O; R 3 ═CH 3 ; R 4 ═CH 2 (CH 2 ) m —NH—C(═O)-spermine; m=1-6, y=0 where B is selected from the group —O(CH 2 ) i , —S(CH 2 ) i , —S—S(CH 2 ) i , —SO 2 —(CH 2 ) i , i=1-5; or   W 1 ═W 2 ═—CH 2 —B—CH 2 —N(R 3 R 4 ), R 1 ═R 2 ═C 8 -C 22  alkyl, X═O; R 3 ═CH 3 ; R 4 ═CH 2 (CH 2 ) m —NH—(C═O)-amino acid side chain; m=1-6, y=0 where B is selected from the group —O(CH 2 ) i , —S(CH 2 ) i , —S—S(CH 2 ) i , —SO 2 —(CH 2 ) i , i=1-5; or   W 1 ═W 2 ═—CH 2 —B—CH 2 —N(R 3 R 4 ), R 1 ═R 2 ═C 8 -C 22  alkyl, X═O; R 3 ═CH 3 ; R 4 ═(CH 2 ) m (CH—OH)(CH 2 )m-NH 2 , y=0 where B is selected from the group —O(CH 2 ) i , —S(CH 2 ) i , —S—S(CH 2 ) i , —SO 2 —(CH 2 ) i , i=1-5; or   W 1 ═W 2 ═—CH 2 —B—CH 2 —N(R 3 R 4 ), R 1 ═R 2 ═C 8 -C 22  alkyl, X═O; R 3 ═CH 3 ; R 4 ═CH 2 (CH 2 )n-NH 2 , y=0 where B is selected from the group —O(CH 2 ) i , —S(CH 2 ) i , —S—S(CH 2 ) i , —SO 2 —(CH 2 ) i , i=1-5; or   W 1 ═H W 2 ═—CH 2 —B—CH 2 —N(R 3 R 4 )CH 2 —R 5 ; R 3 ═R 4 ═CH 3 ; R 5 ═H, y=0 where B is selected from the group —O(CH 2 ) i , —S(CH 2 ) i , —S—S(CH 2 ) i , —SO 2 —(CH 2 ) i , i=1-5; or   W 1 ═H; W 2 ═CH 2 —O—P(═O)(OMe)-O—CH 2 CH 2 —NH—C(═O)-spermine, y=0; or   W 1 ═H; W 2 ═CH 2 —O—P(═O)(OMe)-O—CH 2 CH 2 —NH—C(═O)-amino acid side chain, y=0; or   W 1 ═H; W 2 ═CH 2 —O—P(═O)(OMe)-O—CH 2 CH 2 —N + (CH 3 ) 3 C 1 , y=0; or   W 1 ═H; W 2 ═CH 2 —O—P(═O)(O)—O—CH 2 CH 2 —NH—C(═O)-spermine, y=0; or   Z═(CH 2 ) q , W 1 ═W 2 ═—CH 2 —B—CH 2 —N(R 3 R 4 ), R 1 ═R 2 ═C 8 -C 22  alkyl, X═O; R 3 ═CH 3 , R 4 ═—C(═O)—C(NH 2 )—CH 2 —(CH 2 )n-NH 2 , n=0-6,q=1-3, y=1 where B is selected from the group —O(CH 2 ) i , —S(CH 2 ) i , —S—S(CH 2 ) i , —SO 2 —(CH 2 ) i , i=1-5; or   Z═(CH 2 ) q , W 1 ═W 2 ═—CH 2 —B—CH 2 —N(R 3 R 4 ), R 1 ═R 2 ═C 8 -C 22  alkyl, X═O; R 3 ═CH 3 , R 4 ═—C(═O)—C(NH 2 )—CH 2 —(CH 2 ) n —OH, n=0-6,q=1-3, y=1 where B is selected from the group —O(CH 2 ) i , —S(CH 2 ) i , —S—S(CH 2 ) i , —SO 2 —(CH 2 ) i , i=1-5; or   Z═(CH 2 ) q , W1═W 2 ═—CH 2 —B—CH 2 —N(R 3 R 4 ), R 1 ═R 2 ═C 8 -C 22  alkyl, X═O; R 3 ═CH 3 , R 4 ═—C(═O)—C(OH)—CH 2 —(CH 2 ) n —NH 2 , n=0-6, q=1-3, y=1 where B is selected from the group —O(CH 2 ) i , —S(CH 2 ) i , —S—S(CH 2 ) i , —SO 2 —(CH 2 ) i , i=1-5; or   Z═(CH 2 ) q , W 1 ═W 2 ═—CH 2 —B—CH 2 —N(R 3 R 4 ), R 1 ═R 2 ═C 8 -C 22  alkyl, X═O; R 3 ═CH 3 , R 4 ═—C(═O)—C(NH-iso-butyl)-CH 2 —(CH 2 ) n —O-iso-butyl, n=0-6,y=1 where B is selected from the group —O(CH 2 ) i , —S(CH 2 ) i , —S—S(CH 2 ) i , —SO 2 —(CH 2 ) i , i=1-5; or   Z═(CH 2 ) q , W 1 ═W 2 ═—CH 2 —B—CH 2 —N(R 3 R 4 ), R 1 ═R 2 ═C 8 -C 22  alkyl, X═O; R 3 ═CH 3 , R 4 ═—C(═O)—C(NH 2 )—CH 2 —(CH 2 ) n —C(═NH)—NH 2 , n=0-6, q=1-3, y=1 where B is selected from the group —O(CH 2 ) i , —S(CH 2 ) i , —S—S(CH 2 ) i , —SO 2 —(CH 2 ) i , i=1-5; or   Z═(CH 2 ) q , W 1 ═W 2 ═—CH 2 —B—CH 2 —N(R 3 R 4 ), R 1 ═R 2 ═C 8 -C 22  alkyl, X═O; R 3 ═CH 3 , R 4 ═—C(═O)—C(NH 2 )—CH 2 —(CH 2 ) n -His, n=0-6, q=1-3, y=1 where B is selected from the group —O(CH 2 ) i , —S(CH 2 ) i , —S—S(CH 2 ) i , —SO 2 —(CH 2 ) i , i=1-5; or   Z═(CH 2 ) q W 1 ═W 2 ═—CH 2 —B—CH 2 —N(R 3 R 4 ), R 1 ═R 2 ═C 8 -C 22  alkyl, X═O; R 3 ═CH 3 , R 4 ═—C(═O)-spermine, q=1-3, y=1 where B is selected from the group —O(CH 2 ) i , —S(CH 2 ) i , —S—S(CH 2 ) i , —SO 2 —(CH 2 ) i , i=1-5; or   Z═(CH 2 ) q , W 1 ═W 2 ═—CH 2 —B—CH 2 —N(R 3 R 4 ), R 1 ═R 2 ═C 8 -C 22  alkyl, X═O; R 3 ═CH 3 ; R 4 ═H, q=1-3, y=1 where B is selected from the group —O(CH 2 ) i , —S(CH 2 ) i , —S—S(CH 2 ) i , —SO 2 —(CH 2 ) i , i=1-5; or   Z═(CH 2 ) q W 1 ═W 2 ═—CH 2 —B—CH 2 —N(R 3 R 4 ), R 1 ═R 2 ═C 8 -C 22  alkyl, X═O; R 3 ═CH 3 ; R 4 ═CH 2 (CH 2 ) m —NH 2 ; m=1-6, q=1-3, y=1 where B is selected from the group —O(CH 2 ) i , —S(CH 2 ) i , —S—S(CH 2 ) i , —SO 2 —(CH 2 ) i , i=1-5; or   Z═(CH 2 ) q , W 1 ═W 2 ═—CH 2 —B—CH 2 —N(R 3 R 4 ), R 1 ═R 2 ═C 8 -C 22  alkyl, X═O; R 3 ═CH 3 ; R 4 ═CH 2 (CH 2 ) m —NH—C(═O)-spermine; m=1-6, q=1-3, y=1 where B is selected from the group —O(CH 2 ) i , —S(CH 2 ) i , —S—S(CH 2 ) i , —SO 2 —(CH 2 ) i , i=1-5; or   Z═(CH 2 ) q , W 1 ═W 2 ═—CH 2 —B—CH 2 —N(R 3 R 4 ), R 1 ═R 2 ═C 8 -C 22  alkyl, X═O; R 3 ═CH 3 ; R 4 ═CH 2 (CH 2 ) m —NH—(C═O)-amino acid side chain; m=1-6, q=1-3, y=1 where B is selected from the group —O(CH 2 ) i , —S(CH 2 ) i , —S—S(CH 2 ) i , —SO 2 —(CH 2 ) i , i=1-5; or   Z═(CH 2 ) q , W 1 ═W 2 ═—CH 2 —B—CH 2 —N(R 3 R 4 ), R 1 ═R 2 ═C 8 -C 22  alkyl, X═O; R 3 ═CH 3 ; R 4 ═(CH 2 ) m (CH—OH)(CH 2 ) m —NH 2 , q=1-3, y=1 where B is selected from the group —O(CH 2 ) i , —S(CH 2 ) i , —S—S(CH 2 ) i , —SO 2 —(CH 2 ) i , i=1-5; or   Z═(CH 2 ) q , W 1 ═W 2 ═—CH 2 —B—CH 2 —N(R 3 R 4 ), R 1 ═R 2 ═C 8 -C 22  alkyl, X═O; R 3 ═CH 3 ; R 4 ═CH 2 (CH 2 ) n —NH 2 , q=1-3, y=1 where B is selected from the group —O(CH 2 ) i , —S(CH 2 ) i , —S—S(CH 2 ) i , —SO 2 —(CH 2 ) i , i=1-5; or   Z═(CH 2 ) q —N(R 3 )—(CH 2 ) q , W 1 ═W 2 ═—CH 2 —B—CH 2 —N(R 3 R 4 ), R 1 ═R 2 ═C 8 -C 22  alkyl, X═O; R 3 ═CH 3 , R 4 ═—C(═O)—C(NH 2 )—CH 2 —(CH 2 )n-NH 2 , n=0-6, q=1-3, y=1 where B is selected from the group —O(CH 2 ) i , —S(CH 2 ) i , —S—S(CH 2 ) i , —SO 2 —(CH 2 ) i , i=1-5; or   Z═(CH 2 ) p —N(R 3 )—(CH 2 ) p , W 1 ═W 2 ═—CH 2 —B—CH 2 —N(R 3 R 4 ), R 1 ═R 2 ═C 8 -C 22  alkyl, X═O; R 3 ═CH 3 , R 4 ═—C(═O)—C(NH 2 )—CH 2 —(CH 2 )n-OH, n=0-6, q=1-3, y=1 where B is selected from the group —O(CH 2 ) i , —S(CH 2 ) i , —S—S(CH 2 ) i , —SO 2 —(CH 2 ) i , i=1-5; or   Z═(CH 2 ) q —N(R 3 )—(CH 2 ) q , W1═W 2 ═—CH 2 —B—CH 2 —N(R 3 R 4 ), R 1 ═R 2 ═C 8 -C 22  alkyl, X═O; R 3 ═CH 3 , R 4 ═—C(═O)—C(OH)—CH 2 —(CH 2 )n-NH 2 , n=0-6, q=1-3, y=1 where B is selected from the group —O(CH 2 ) i , —S(CH 2 ) i , —S—S(CH 2 ) i , —SO 2 —(CH 2 ) i , i=1-5; or   Z═(CH 2 ) q —N(R 3 )—(CH 2 ) q , W 1 ═W 2 ═—CH 2 —B—CH 2 —N(R 3 R 4 ), R 1 ═R 2 ═C 8 -C 22  alkyl, X═O; R 3 ═CH 3 , R 4 ═—C(═O)—C(NH-iso-butyl)-CH 2 —(CH 2 )n-O-iso-butyl, n=0-6,y=1l where B is selected from the group —O(CH 2 ) i , —S(CH 2 ) i , —S—S(CH 2 ) i , —SO 2 —(CH 2 ) i , i=1-5; or   Z═(CH 2 ) q —N(R 3 )—(CH 2 ) q , W 1 ═W 2 ═—CH 2 —B—CH 2 —N(R 3 R 4 ), R 1 ═R 2 ═C 8 -C 22  alkyl, X═O; R 3 ═CH 3 , R 4 ═—C(═O)—C(NH 2 )—CH 2 —(CH 2 )n-C(═NH)—NH 2 , n=0-6, q=1-3, y=1 where B is selected from the group —O(CH 2 ) i , —S(CH 2 ) i , —S—S(CH 2 ) i , —SO 2 —(CH 2 ) i , i=1-5; or   Z═(CH 2 ) q —N(R 3 )—(CH 2 ) q , W 1 ═W 2 ═—CH 2 —B—CH 2 —N(R 3 R 4 ), R 1 ═R 2 ═C 8 -C 22  alkyl, X═O; R 3 ═CH 3 , R 4 ═—C(═O)—C(NH 2 )—CH 2 —(CH 2 )n-His, n=0-6, q=1-3, y=1 where B is selected from the group —O(CH 2 ) i , —S(CH 2 ) i , —S—S(CH 2 ) i , —SO 2 —(CH 2 ) i , i=1-5; or   Z═(CH 2 ) q —N(R 3 )—(CH 2 ) q , W 1 ═W 2 ═—CH 2 —B—CH 2 —N(R 3 R 4 ), R 1 ═R 2 ═C 8 -C 22  alkyl, X═O; R 3 ═CH 3 , R 4 ═—C(═O)-spermine, q=1-3, y=1 where B is selected from the group —O(CH 2 ) i , —S(CH 2 ) i , —S—S(CH 2 ) i , —SO 2 —(CH 2 ) i , i=1-5; or   Z═(CH 2 )q-N(R 3 )—(CH 2 )q, W 1 ═W 2 ═—CH 2 —B—CH 2 —N(R 3 R 4 ), R 1 ═R 2 ═C 8 -C 22  alkyl, X═O; R 3 ═CH 3 ; R 4 ═H, q=1-3, y=1 where B is selected from the group —O(CH 2 ) i , —S(CH 2 ) i , —S—S(CH 2 ) i , —SO 2 —(CH 2 ) i , i=1-5; or   Z═(CH 2 ) q —N(R 3 )—(CH 2 ) q , W 1 ═W 2 ═—CH 2 —B—CH 2 —N(R 3 R 4 ), R 1 ═R 2 ═C 8 -C 22  alkyl, X═O; R 3 ═CH 3 ; R 4 ═CH 2 (CH 2 ) m —NH 2 ; m=1-6, q=1-3, y=1 where B is selected from the group —O(CH 2 ) i , —S(CH 2 ) i , —S—S(CH 2 ) i , —SO 2 —(CH 2 ) i , i=1-5; or   Z═(CH 2 ) q —N(R 3 )—(CH 2 ) q , W 1 ═W 2 ═—CH 2 —B—CH 2 —N(R 3 R 4 ), R 1 ═R 2 ═C 8 -C 22  alkyl, X═O; R 3 ═CH 3 ; R 4 ═CH 2 (CH 2 ) m —NH—C(═O)-spermine; m=1-6, q=1-3, y=1 where B is selected from the group —O(CH 2 ) i , —S(CH 2 ) i , —S—S(CH 2 ) i , —SO2-(CH 2 ) i , i=1-5; or   Z═(CH 2 ) q —N(R 3 )—(CH 2 ) q , W 1 ═W 2 ═—CH 2 —B—CH 2 —N(R 3 R 4 ), R 1 ═R 2 ═C 8 -C 22  alkyl, X═O; R 3 ═CH 3 ; R 4 ═CH 2 (CH 2 ) m —NH—(C═O)-amino acid side chain; m=1-6, q=1-3, y=1 where B is selected from the group —O(CH 2 ) i , —S(CH 2 ) i , —S—S(CH 2 ) i , —SO 2 —(CH 2 ) i , i=1-5; or   Z═(CH 2 ) q —N(R 3 )—(CH 2 ) q , W 1 ═W 2 ═—CH 2 —B—CH 2 —N(R 3 R 4 ), R 1 ═R 2 ═C 8 -C 22  alkyl, X═O; R 3 ═CH 3 ; R 4 ═(CH 2 ) m (CH—OH)(CH 2 ) m —NH 2 , q=1-3, y=1 where B is selected from the group —O(CH 2 ) i , —S(CH 2 ) i , —S—S(CH 2 ) i , —SO 2 —(CH 2 ) i , i=1-5; or   Z═(CH 2 ) q —N(R 3 )—(CH 2 ) q , W 1 ═W 2 ═—CH 2 —B—CH 2 —N(R 3 R 4 ), R 1 ═R 2 ═C 8 -C 22  alkyl, X═O; R 3 ═CH 3 ; R 4 ═CH 2 (CH 2 ) n —NH 2 , q=1-3, y=1 where B is selected from the group —O(CH 2 ) i , —S(CH 2 ) i , —S—S(CH 2 ) i , —SO 2 —(CH 2 ) i , i=1-5;   Z═(CH 2 ) q —S—S—(CH 2 ) q , W 1 ═W 2 ═—CH 2 —B—CH 2 —N(R 3 R 4 ), R 1 ═R 2 ═C 8 -C 22  alkyl, X═O; R 3 ═CH 3 , R 4 ═—C(═O)—C(NH 2 )—CH 2 —(CH 2 ) n —NH 2 , n=0-6, q=1-3, y=1 where B is selected from the group —O(CH 2 ) i , —S(CH 2 ) i , —S—S(CH 2 ) i , —SO 2 —(CH 2 ) i , i=1-5; or   Z═(CH 2 ) q —S—S—(CH 2 ) q , W 1 ═W 2 ═—CH 2 —B—CH 2 —N(R 3 R 4 ), R 1 ═R 2 ═C 8 -C 22  alkyl, X═O; R 3 ═CH 3 , R 4 ═—C(═O)—C(NH 2 )—CH 2 —(CH 2 ) n —OH, n=0-6, q=1-3, y=11 where B is selected from the group —O(CH 2 ) i , —S(CH 2 ) i , —S—S(CH 2 ) i , —SO 2 —(CH 2 ) i , i=1-5; or   Z═(CH 2 ) q —S—S—(CH 2 ) q , W1═W 2 ═—CH 2 —B—CH 2 —N(R 3 R 4 ), R 1 ═R 2 ═C 8 -C 22  alkyl, X═O; R 3 ═CH 3 , R 4 ═—C(═O)—C(OH)—CH 2 —(CH 2 ) n —NH 2 , n=0-6, q=1-3, y=11 where B is selected from the group —O(CH 2 ) i , —S(CH 2 ) i , —S—S(CH 2 ) i , —SO 2 —(CH 2 ) i , i=1-5; or   Z═(CH 2 ) q —S—S—(CH 2 ) q , W 1 ═W 2 ═—CH 2 —B—CH 2 —N(R 3 R 4 ), R 1 ═R 2 ═C 8 -C 22  alkyl, X═O; R 3 ═CH 3 , R 4 ═—C(═O)—C(NH-iso-butyl)-CH 2 —(CH 2 ) n —O-iso-butyl, n=0-6,y=1 where B is selected from the group —O(CH 2 ) i , —S(CH 2 ) i , —S—S(CH 2 ) i , —SO 2 —(CH 2 ) i , i=1-5; or   Z═(CH 2 ) q —S—S—(CH 2 ) q , W 1 ═W 2 ═—CH 2 —B—CH 2 —N(R 3 R 4 ), R 1 ═R 2 ═C 8 -C 22  alkyl, X═O; R 3 ═CH 3 , R 4 ═—C(═O)—C(NH 2 )—CH 2 —(CH 2 ) n —C(═NH)—NH 2 , n=0-6, q=1-3, y=1 where B is selected from the group —O(CH 2 ) i , —S(CH 2 ) i , —S—S(CH 2 ) i , —SO 2 —(CH 2 ) i , i=1-5; or   Z═(CH 2 ) q —S—S—(CH 2 ) q , W 1 ═W 2 ═—CH 2 —B—CH 2 —N(R 3 R 4 ), R 1 ═R 2 ═C 8 -C 22  alkyl, X═O; R 3 ═CH 3 , R 4 ═—C(═O)—C(NH 2 )—CH 2 —(CH 2 ) n -His, n=0-6, q=1-3, y=11 where B is selected from the group —O(CH 2 ) i , —S(CH 2 ) i , —S—S(CH 2 ) i , —SO 2 —(CH 2 ) i , i=1-5; or   Z═(CH 2 ) q —S—S—(CH 2 ) q , W 1 ═W 2 ═—CH 2 —B—CH 2 —N(R 3 R 4 ), R 1 ═R 2 ═C 8 -C 22  alkyl, X═O; R 3 ═CH 3 , R 4 ═—C(═O)-spermine, q=1-3, y=1 where B is selected from the group —O(CH 2 ) i , —S(CH 2 ) i , —S—S(CH 2 ) i , —SO 2 —(CH 2 ) i , i=1-5; or   Z═(CH 2 ) q —S—S—(CH 2 ) q , W 1 ═W 2 ═—CH 2 —B—CH 2 —N(R 3 R 4 ), R 1 ═R 2 ═C 8 -C 22  alkyl, X═O; R 3 ═CH 3 ; R 4 ═H, q=1-3, y=1 where B is selected from the group —O(CH 2 ) i , —S(CH 2 ) i , —S—S(CH 2 ) i , —SO 2 —(CH 2 ) i , i=1-5; or   Z═(CH 2 ) q —S—S—(CH 2 ) q , W 1 ═W 2 ═—CH 2 —B—CH 2 —N(R 3 R 4 ), R 1 ═R 2 ═C 8 -C 22  alkyl, X═O; R 3 ═CH 3 ; R 4 ═CH 2 (CH 2 ) m —NH 2 ; m=1-6, q=1-3, y=1 where B is selected from the group —O(CH 2 ) i , —S(CH 2 ) i , —S—S(CH 2 ) i , —SO 2 —(CH 2 ) i , i=1-5; or   Z═(CH 2 ) q —S—S—(CH 2 ) q , W 1 ═W 2 ═—CH 2 —B—CH 2 —N(R 3 R 4 ), R 1 ═R 2 ═C 8 -C 22  alkyl, X═O; R 3 ═CH 3 ; R 4 ═CH 2 (CH 2 ) m —NH—C(═O)-spermine; m=1-6, q=1-3, y=1 where B is selected from the group —O(CH 2 ) i , —S(CH 2 ) i , —S—S(CH 2 ) i , —SO 2 —(CH 2 ) i , i=1-5; or   Z═(CH 2 ) q —S—S—(CH 2 ) q , W 1 ═W 2 ═—CH 2 —B—CH 2 —N(R 3 R 4 ), R 1 ═R 2 ═C 8 -C 22  alkyl, X═O; R 3 ═CH 3 ; R 4 ═CH 2 (CH 2 ) m —NH—(C═O)-amino acid side chain; m=1-6, q=1-3, y=1 where B is selected from the group —O(CH 2 ) i , —S(CH 2 ) i , —S—S(CH 2 ) i , —SO 2 —(CH 2 ) i , i=1-5; or   Z═(CH 2 ) q —S—S—(CH 2 ) q , W 1 ═W 2 ═—CH 2 —B—CH 2 —N(R 3 R 4 ), R 1 ═R 2 ═C 8 -C 22  alkyl, X═O; R 3 ═CH 3 ; R 4 ═(CH 2 ) m (CH—OH)(CH 2 ) m —NH 2 , q=1-3, y=1 where B is selected from the group —O(CH 2 ) i , —S(CH 2 ) i , —S—S(CH 2 ) i , —SO 2 —(CH 2 ) i , i=1-5; or   Z═(CH 2 ) q —S—S—(CH 2 ) q , W 1 ═W 2 ═—CH 2 —B—CH 2 —N(R 3 R 4 ), R 1 ═R 2 ═C 8 -C 22  alkyl, X═O; R 3 ═CH 3 ; R 4 ═CH 2 (CH 2 ) n —NH 2 , q=1-3, y=1 where B is selected from the group —O(CH 2 ) i , —S(CH 2 ) i , —S—S(CH 2 ) i , —SO 2 —(CH 2 ) i , i=1-5; or   Z═(CH 2 ) q W 1 ═W 2 ═—CH 2 —B—CH 2 —N(R 3 R 4 ), R 1 ═R 2 ═C 8 -C 22  alkyl, X═O; R 3 ═CH 3 ; R 4 ═COCH 2 (OCH 2 CH 2 ) m —NH 2 ; m=1-6, q=1-3, y=1 where B is selected from the group —O(CH 2 ) i , —S(CH 2 ) i , —S—S(CH 2 ) i , —SO 2 —(CH 2 ) i , i=1-5; or   Z═(CH 2 ) q —S—S—(CH 2 ) q , W 1 ═W 2 ═—CH 2 —B—CH 2 —N(R 3 R 4 ), R 1 ═R 2 ═C 8 -C 22  alkyl, X═O; R 3 ═CH 3 ; R 4 ═COCH 2 (OCH 2 CH 2 ) m —NH 2 ; m=1-6, q=1-3, y=1 where B is selected from the group —O(CH 2 ) i , —S(CH 2 ) i , —S—S(CH 2 ) i , —SO 2 —(CH 2 ) i , i=1-5; or   Z═(CH 2 ) q —N(R 3 )—(CH 2 ) q , W 1 ═W 2 ═—CH 2 —B—CH 2 —N(R 3 R 4 ), R 1 ═R 2 ═C 8 -C 22  alkyl, X═O; R 3 ═CH 3 ; R 4 ═COCH 2 (OCH 2 CH 2 ) m —NH 2 ; m=1-6, q=1-3, y=1 where B is selected from the group —O(CH 2 ) i , —S(CH 2 ) i , —S—S(CH 2 ) i , —SO 2 —(CH 2 ) i , i=1-5; or   Z═(CH 2 ) q —O—(CH 2 ) q , W 1 ═W 2 ═CH 2 —N(R 3 R 4 ), R 1 ═R 2 ═C 8 -C 22  alkyl, X═O; R 3 ═CH 3 , R 4 ═—C(═O)—C(NH 2 )—CH 2 —(CH 2 ) n —NH 2 , n=0-6, q=1-3, y=1; or   Z═(CH 2 ) q —O—(CH 2 ) q , W 1 ═W 2 ═CH 2 —N(R 3 R 4 ), R 1 ═R 2 ═C 8 -C 22  alkyl, X═O; R 3 ═CH 3 , R 4 ═—C(═O)—C(NH 2 )—CH 2 —(CH 2 ) n —OH, n=0-6, q=1-3, y=1; or   Z═(CH 2 ) q —O—(CH 2 ) q , W1═W 2 ═CH 2 —N(R 3 R 4 ), R 1 ═R 2 ═C 8 -C 22  alkyl, X═O; R 3 ═CH 3 , R 4 ═—C(═O)—C(OH)—CH 2 —(CH 2 ) n —NH 2 , n=0-6, q=1-3, y=1; or   Z═(CH 2 ) q —O—(CH 2 ) q , W 1 ═W 2 ═CH 2 —N(R 3 R 4 ), R 1 ═R 2 ═C 8 -C 22  alkyl, X═O; R 3 ═CH 3 , R 4 ═—C(═O)—C(NH-iso-butyl)-CH 2 —(CH 2 ) n —O-iso-butyl, n=0-6,y=1; or   Z═(CH 2 ) q —O—(CH 2 ) q , W 1 ═W 2 ═CH 2 —N(R 3 R 4 ), R 1 ═R 2 ═C 8 -C 22  alkyl, X═O; R 3 ═CH 3 , R 4 ═—C(═O)—C(NH 2 )—CH 2 —(CH 2 ) n —C(═NH)—NH 2 , n=0-6, q=1-3, y=1; or   Z═(CH 2 ) q —O—(CH 2 ) q , W 1 ═W 2 ═CH 2 —N(R 3 R 4 ), R 1 ═R 2 ═C 8 -C 22  alkyl, X═O; R 3 ═CH 3 , R 4 ═—C(═O)—C(NH 2 )—CH 2 —(CH 2 ) n -His, n=0-6, q=1-3, y=1; or   Z═(CH 2 ) q —O—(CH 2 ) q , W 1 ═W 2 ═CH 2 —N(R 3 R 4 ), R 1 ═R 2 ═C 8 -C 22  alkyl, X═O; R 3 ═CH 3 , R 4 ═—C(═O)-spermine, q=1-3, y=1; or   Z═(CH 2 ) q —O—(CH 2 ) q , W 1 ═W 2 ═CH 2 —N(R 3 R 4 ), R 1 ═R 2 ═C 8 -C 22  alkyl, X═O; R 3 ═CH 3 ; R 4 ═H, q=1-3, y=1; or   Z═(CH 2 ) q —O—(CH 2 ) q , W 1 ═W 2 ═CH 2 —N(R 3 R 4 ), R 1 ═R 2 ═C 8 -C 22  alkyl, X═O; R 3 ═CH 3 ; R 4 ═CH 2 (CH 2 ) m —NH 2 ; m=1-6, q=1-3, y=1; or   Z═(CH 2 ) q —O—(CH 2 ) q , W 1 ═W 2 ═CH 2 —N(R 3 R 4 ), R 1 ═R 2 ═C 8 -C 22  alkyl, X═O; R 3 ═CH 3 ; R 4 ═CH 2 (CH 2 ) m —NH—C(═O)-spermine; m=1-6, q=1-3, y=1; or   Z═(CH 2 ) q —O—(CH 2 ) q , W 1 ═W 2 ═CH 2 —N(R 3 R 4 ), R 1 ═R 2 ═C 8 -C 22  alkyl, X═O; R 3 ═CH 3 ; R 4 ═CH 2 (CH 2 ) m —NH—(C═O)-amino acid side chain; m=1-6, q=1-3, y=1; or   Z═(CH 2 ) q —O—(CH 2 ) q , W 1 ═W 2 ═CH 2 —N(R 3 R 4 ), R 1 ═R 2 ═C 8 -C 22  alkyl, X═O; R 3 ═CH 3 ; R 4 ═(CH 2 ) m (CH—OH)(CH 2 ) m —NH 2 , q=1-3, y=1; or   Z═(CH 2 ) q —O—(CH 2 ) q , W 1 ═W 2 ═CH 2 —N(R 3 R 4 ), R 1 ═R 2 ═C 8 -C 22  alkyl, X═O; R 3 ═CH 3 ; R 4 ═CH 2 (CH 2 ) n —NH 2 , q=1-3, y=1; or   Z═(CH 2 ) q —O—(CH 2 ) q , W 1 ═W 2 ═—CH 2 —B—CH 2 —N(R 3 R 4 ), R 1 ═R 2 ═C 8 -C 22  alkyl, X═O; R 3 ═CH 3 , R 4 ═—C(═O)—C(NH 2 )—CH 2 —(CH 2 ) n —NH 2 , n=0-6, q=1-3, y=1 where B is selected from the group —O(CH 2 ) i , —S(CH 2 ) i , —S—S(CH 2 ) i , —SO 2 —(CH 2 ) i , i=1-5; or   Z═(CH 2 ) q —O—(CH 2 ) q , W 1 ═W 2 ═—CH 2 —B—CH 2 —N(R 3 R 4 ), R 1 ═R 2 ═C 8 -C 22  alkyl, X═O; R 3 ═CH 3 , R 4 ═—C(═O)—C(NH 2 )—CH 2 —(CH 2 ) n —OH, n=0-6, q=1-3, y=1 where B is selected from the group —O(CH 2 ) i , —S(CH 2 ) i , —S—S(CH 2 ) i , —SO 2 —(CH 2 ) i , i=1-5; or   Z═(CH 2 ) q —O—(CH 2 ) q , W1═W 2 ═—CH 2 —B—CH 2 —N(R 3 R 4 ), R 1 ═R 2 ═C 8 -C 22  alkyl, X═O; R 3 ═CH 3 , R 4 ═—C(═O)—C(OH)—CH 2 —(CH 2 ) n —NH 2 , n=0-6, q=1-3, y=1 where B is selected from the group —O(CH 2 ) i , —S(CH 2 ) i , —S—S(CH 2 ) i , —SO 2 —(CH 2 ) i , i=1-5; or   Z═(CH 2 ) q —O—(CH 2 ) q , W 1 ═W 2 ═—CH 2 —B—CH 2 —N(R 3 R 4 ), R 1 ═R 2 ═C 8 -C 22  alkyl, X═O; R 3 ═CH 3 , R 4 ═—C(═O)—C(NH-iso-butyl)-CH 2 —(CH 2 ) n —O-iso-butyl, n=0-6,y=1 where B is selected from the group —O(CH 2 ) i , —S(CH 2 ) i , —S—S(CH 2 ) i , —SO 2 —(CH 2 ) i , i=1-5; or   Z═(CH 2 ) q —O—(CH 2 ) q , W 1 ═W 2 ═—CH 2 —B—CH 2 —N(R 3 R 4 ), R 1 ═R 2 ═C 8 -C 22  alkyl, X═O; R 3 ═CH 3 , R 4 ═—C(═O)—C(NH 2 )—CH 2 —(CH 2 ) n —C(═NH)—NH 2 , n=0-6, q=1-3, y=1 where B is selected from the group —O(CH 2 ) i , —S(CH 2 ) i , —S—S(CH 2 ) i , —SO 2 —(CH 2 ) i , i=1-5; or   Z═(CH 2 ) q —O—(CH 2 ) q , W 1 ═W 2 ═—CH 2 —B—CH 2 —N(R 3 R 4 ), R 1 ═R 2 ═C 8 -C 22  alkyl, X═O; R 3 ═CH 3 , R 4 ═—C(═O)—C(NH 2 )—CH 2 —(CH 2 ) n -His, n=0-6, q=1-3, y=1 where B is selected from the group —O(CH 2 ) i , —S(CH 2 ) i , —S—S(CH 2 ) i , —SO 2 —(CH 2 ) i , i=1-5; or   Z═(CH 2 ) q —O—(CH 2 ) q , W 1 ═W 2 ═—CH 2 —B—CH 2 —N(R 3 R 4 ), R 1 ═R 2 ═C 8 -C 22  alkyl, X═O; R 3 ═CH 3 , R 4 ═—C(═O)-spermine, q=1-3, y=1 where B is selected from the group —O(CH 2 ) i , —S(CH 2 ) i , —S—S(CH 2 ) i , —SO 2 —(CH 2 ) i , i=1-5; or   Z═(CH 2 ) q —O—(CH 2 ) q , W 1 ═W 2 ═—CH 2 —B—CH 2 —N(R 3 R 4 ), R 1 ═R 2 ═C 8 -C 22  alkyl, X═O; R 3 ═CH 3 ; R 4 ═H, q=1-3, y=1 where B is selected from the group —O(CH 2 ) i , —S(CH 2 ) i , —S—S(CH 2 ) i , —SO 2 —(CH 2 ) i , i=1-5; or   Z═(CH 2 ) q —O—(CH 2 ) q , W 1 ═W 2 ═—CH 2 —B—CH 2 —N(R 3 R 4 ), R 1 ═R 2 ═C 8 -C 22  alkyl, X═O; R 3 ═CH 3 ; R 4 ═CH 2 (CH 2 ) m —NH 2 ; m=1-6, q=1-3, y=1 where B is selected from the group —O(CH 2 ) i , —S(CH 2 ) i , —S—S(CH 2 ) i , —SO 2 —(CH 2 ) i , i=1-5; or   Z═(CH 2 ) q —O—(CH 2 ) q , W 1 ═W 2 ═—CH 2 —B—CH 2 —N(R 3 R 4 ), R 1 ═R 2 ═C 8 -C 22  alkyl, X═O; R 3 ═CH 3 ; R 4 ═CH 2 (CH 2 ) m —NH—C(═O)-spermine; m=1-6, q=1-3, y=1 where B is selected from the group —O(CH 2 ) i , —S(CH 2 ) i , —S—S(CH 2 ) i , —SO 2 —(CH 2 ) i , i=1-5; or   Z═(CH 2 ) q —O—(CH 2 ) q , W 1 ═W 2 ═—CH 2 —B—CH 2 —N(R 3 R 4 ), R 1 ═R 2 ═C 8 -C 22  alkyl, X═O; R 3 ═CH 3 ; R 4 ═CH 2 (CH 2 ) m —NH—(C═O)-amino acid side chain; m=1-6, q=1-3, y=1 where B is selected from the group —O(CH 2 ) i , —S(CH 2 ) i , —S—S(CH 2 ) i , —SO 2 —(CH 2 ) i , i=1-5.       

     In some embodiments, the compound of Formula I is a compound where:
         R 1 ═R 2 ═C 8 -C 22  alkyl, X═O, W 1 ═H; W 2 ═CH 2 —O—P(═O)(OMe)-O—CH 2 CH 2 —NH—C(═O)-spermine, y=0;   W 1 ═W 2 ═CH 2 —N(R 3 R 4 ), R 1 ═R 2 ═C 8 -C 22  alkyl, X═O; R 3 ═CH 3 , R 4 ═—C(═O)—C(NH 2 )—CH 2 —(CH 2 ) n —C(═NH)—NH 2 , n=0-6, y=0;   W 1 ═W 2 ═CH 2 —N(R 3 R 4 ), R 2 ═C 8 -C 22  alkyl, X═O; R 3 ═CH 3 , R 4 ═—C(═O)-spermine, y=0;   W 1 ═W 2 ═CH 2 —N(R 3 R 4 ), R 2 ═C 8 -C 22  alkyl, X═O; R 3 ═CH 3 , R 4 ═—C(═O)—C(NH 2 )—CH 2 —(CH 2 ) n —NH 2 , n=0-6, y=0; or   W 1 ═W 2 ═CH 2 —N(R 3 R 4 ), R 2 ═C 8 -C 22  alkyl, X═O; R 3 ═CH 3 ; R 4 ═(CH 2 ) m (CH—OH)(CH 2 ) m —NH 2 , y=0.       

     In other embodiments, the compound of Formula I is a compound where: R 1 ═R 2 ═C 8 -C 22  alkyl, X═O, W 1 ═H; W 2 ═CH 2 —O—P(═O)(OMe)-O—CH 2 CH 2 —NH—C(═O)-spermine, y=0; or
         W 1 ═W 2 ═CH 2 —N(R 3 R 4 ), R 1 ═R 2 ═C 8 -C 22  alkyl, X═O; R 3 ═CH 3 , R 4 ═—C(═O)—C(NH 2 )—CH 2 —(CH 2 ) n —C(═NH)—NH 2 , n=0-6, y=0.       

     In other embodiments, the compound of Formula I is a compound where:
         W 1 ═W 2 ═CH 2 —N(R 3 R 4 ), R 2 ═C 8 -C 22  alkyl, X═O; R 3 ═CH 3 , R 4 ═—C(═O)—C(NH 2 )—CH 2 —(CH 2 )n-NH 2 , n=0-6, y=0; or   W 1 ═W 2 ═CH 2 —N(R 3 R 4 ), R 2 ═C 8 -C 22  alkyl, X═O; R 3 ═CH 3 ; R 4 ═(CH 2 ) m (CH—OH)(CH 2 ) m —NH 2 , y=0.       

     In other embodiments, the compound of Formula I is a compound where:
         R 1 ═R 2 ═C 8 -C 22  alkyl, X═O, W 1 ═H; W 2 ═CH 2 —O—P(═O)(OMe)-O—CH 2 CH 2 —NH—C(═O)-spermine, y=0;   W 1 ═W 2 ═CH 2 —N(R 3 R 4 ), R 1 ═R 2 ═C 8 -C 22  alkyl, X═O; R 3 ═CH 3 , R 4 ═—C(═O)—C(NH 2 )—CH 2 —(CH 2 ) n —C(═NH)—NH 2 , n=0-6, y=0; or   W 1 ═W 2 ═CH 2 —N(R 3 R 4 ), R 2 ═C 8 -C 22  alkyl, X═O; R 3 ═CH 3 , R 4 ═—C(═O)-spermine, y=0.       

     In some embodiments, the transfection agent comprises at least one or more cationic lipid from Formula I and may optionally also contain one or more neutral lipids; DOPE, DPhPE, saturated and unsaturated DPPE, saturated and unsaturated DMPE, cholesterol, DOPC, Lyso-PE (1-acyl-2-hydroxy-sn-glycero-3-phosphoethanolamine), Lyso-PC (1-acyl-3-hydroxy-sn-glycero-3-phosphocholine), 3-alkyloxy-2-hydroxy-1-acetamidopropane, 4-alkyloxy-3-hydroxy-1-acetamidopropane, 5-alkyloxy-4-hydroxy-1-acetamidopropane or 6-alkyloxy-5-hydroxy-1-acetamidopropane. In some embodiments, the alkyloxy in the above list is selected from the group consisting of myristyloxy, myristeleyloxy lauryloxy, palmityloxy, palmitoleyloxy, oleyloxy and streayloxy. In some embodiments transfection agents contains Lyso-phosphatidylcholine, Sphingomyelin, Disaturated phosphatidylcholine, saturated and unsaturated Phosphatidylcholine, Disaturated phosphatidylethanolamine, Phosphatidylethanolamine saturated and unsaturated, Phosphatidylserine, phosphatidylinositol, Lyso-bis phosphatidic acid, Cholesterol, and Diglyceride. Helper Lipids may include, complete exosomes solutions, total extract of lipids isolated from exosomes. 
     In other embodiment, the transfection agent comprises at least one or more cationic lipid of Formula I. In certain embodiments, the transfection agent optionally comprises one or more of cationic lipid, while in other embodiments, the agent optionally comprises one or more neutral lipids or one or more exosomes, or lipid extract for exosomes. In certain embodiments, the transfection agent comprises both one or more cationic lipid and one or more neutral lipid. 
     In some embodiments, the cationic lipid is selected from the group consisting of GeneIn™ (MTI-GlobalStem), TransfeX™ (ATCC), LipofectAmine™ 2000, LipofectAmine 3000, LipofectAmine™, Lipofectin®, DMRIE-C, CellFectin® (Invitrogen), Oligofectamine® (Invitrogen), LipofectAce® (Invitrogen), Fugene® (Promega), Fugene® HD (Promega), Transfectam® (Promega), Tfx-10® (Promega), Tfx-20® (Promega), Tfx-50® (Promega), DNA-In (MTI-GlobalStem), Transfectin™ (BioRad, Hercules, Calif.), SilentFect™ (Bio-Rad), Effectene® (Qiagen, Valencia, Calif.), DC-chol (Avanti Polar Lipids), GenePorter® (Gene Therapy Systems, San Diego, Calif.), DharmaFect 1® (Dharmacon, Lafayette, Colo.), DharmaFect 2® (Dharmacon), DharmaFect 3® (Dharmacon), DharmaFect 4® (Dharmacon), Escort™ III (Sigma, St. Louis, Mo.), Escort™ IV (Sigma), ViaFect™ (Promega), DOTMA, DOTAP, DMRIE, DC-Chol, DDAB, DOSPA, DOSPER, DOGS, TMTPS, TMTOS, TMTLS, TMTMS, TMDOS, N-1-dimethyl-N-1-(2,3-diaoleoyloxypropyl)-2-hydroxypropane-1,3-diamine, N-1-dimethyl-N-1-(2,3-diamyri styloxypropyl)-2-hydroxypropane-1,3-diamine, N-1-dimethyl-N-1-(2,3-diapalmityloxypropyl)-2-hydroxypropane-1,3-diamine, N-1-dimethyl-N-1-(2,3-diaoleoyl-oxypropyl)-2-(3-amino-2-hydroxypropyloxy)propane-1,3-diamine, N-1-dimethyl-N-1-(2,3-diamyristyloxypropyl)-2-(3-amino-2-hydroxypropyloxy)propane-1,3-diamine, N-1-dimethyl-N-1-(2,3-diapalmityloxypropyl)-2-(3-amino-2-hydroxypropyloxy)propane-1,3-diamine, L-spermine-5-carboxyl-3-(DL-1,2-dipalmitoyl-dimethylaminopropyl-β-hydr-oxyethylamine, 3,5-(N,N-di-lysyl)-diaminobenzoyl-glycyl-3-(DL-1,2-dipalmitoyl-di-methylaminopropyl-β-hydroxyethylamine), L-Lysine-bis(O,O′-oleoyl-β-hydroxyethyl)-amide dihydrochloride, L-Lysine-bis-(O,O′-palmitoyl-β-hydroxyethyl)amide dihydrochloride, 1,4-bis[(3-(3-aminopropyl)-alkylamino)-2-hydroxypropyl)piperazine, L-Lysine-bis-(O,O′-myristoyl-β-hydroxyethyl)amide dihydrochloride, L-Omithine-bis-(O,O′-myristoyl-β-hydroxyethyl)amide dihydrochloride, L-Ornithine-bis-(O,O′-oleoyl-β-hydroxyethyl)amide dihydrochloride, 1,4-bis[(3-(3-aminopropyl)-oleylamino)-2-hydroxypropyl]piperazine, L-Omithine-bis-(O,O′-palmitoyl-β-hydroxyethyl)amide dihydrochloride, 1,4,-bis[(3-amino-2-hydroxypropyl)-oleylamino]-butane-2,3-diol, 1,4,-bis[(3-amino-2-hydroxypropyl)-palmitylamino]-butane-2,3-diol, 1,4,-bis[(3-amino-2-hydroxypropyl)-myristylamino]-butane-2,3-diol, 1,4-bis[(3-oleylamino)propyl]piperaz-ine, L-Arginine-bis-(O,O′-oleoyl-β-hydroxyethyl)amide dihydrochloride, bis[(3-(3-aminopropyl)-myristylamino)2-hydroxypropyl]piperazine, L-Arginine-bis-(O,O′-palmitoyl-β-hydroxyethyl)amide dihydrochloride, L-Serine-bis-(O,O′-oleoyl-β-hydroxyethyl)amide dihydrochloride, 1,4-bis[(3-(3-aminopropyl)-palmitylamino)-2-hydroxypropyl]piperazine, Glycine-bis-(O,O′-palmitoyl-β-hydroxyethyl)amide dihydro-chloride, Sarcosine-bis-(O,O′-palmitoyl-β-hydroxyethyl)amide dihydrochloride, L-Histidine-bis-(O,O′-palmitoyl-(3-hydroxyethyl)amide dihydrochloride, cholesteryl-3β-carboxyl-amidoethylenetrimethylammonium iodide, 1,4-bis[(3-myristylamino)propyl]-piperazine, 1-dimethylamino-3-trimethylammonio-DL-2-propyl-cholesteryl carboxylate iodide, cholesteryl-33-carboxyamidoethyleneamine, cholesteryl-3β-oxysuccinamidoethyl-enetrimethylammonium iodide, 1-dimethylamino-3-trimethylammonio-DL-2-propyl-cholesteryl-3β-oxysuccinate iodide, 2-[(2-trimethylammonio)-ethylmethylamino] ethyl-cholesteryl-3β-oxysuccinate iodide, 303[N—(N′,N′-dimethylaminoethane)carbamoyl]-cholesterol, and 303-[N-(polyethyleneimine)-carbamoyl] cholesterol, 1,4-bis[(3-palmitylamino)propyl]piperazine, L-Ornithylglycyl-N-(1-heptadecyloctadecyl)glycin-amide, N 2 ,N 5 -Bis(3-aminopropyl)-L-omithylglycyl-N-(1-heptadecyloctadecyl)glycin-amide, 1,4-bis[(3-(3-amino-2-hydroxypropyl)-alkylamino)-2-hydroxypropyl]piperazine N 2 —[N 2 ,N 5 -Bis(3-aminopropyl)-L-omnithyl]-N,N-dioctadecyl-L-glutamine,N 2 —[N 2 ,N 5 -Bis(aminopropyl)-L-ornithyl]-N—N-dioctadecyl-L-α-glutamine, 1,4-bis[(3-(3-amino-2-hydroxypropyl)-oleylamino)2-hydroxypropyl]piperazine, N 2 —[N 2 ,N 5 -Bis(aminopropyl)-L-ornithyl]-N—N-dioctadecyl-L-α-asparagine, N—[N 2 —[N 2 ,N 5 -Bis[(1,1-dimethylethoxy)-carbonyl]-N 2 ,N 5 -bis[3-[(1,1-dimethylethoxy)carbonyl]aminopropyl]-L-ornithyl-N—N-dioctadecyl-L-glutaminyl]-L-glutamic acid, N 2 —[N 2 ,N 5 -Bis(3-aminopropyl)-L-ornithyl]-N,N-diolyl-L-glutamine, N 2 —[N 2 ,N 5 -Bis(aminopropyl)-L-ornithyl]-N—N-dioleyl-L-α-glutamine,4-bis[(3-(3-amino-2-hydoxypropyl)-myristylamino)-2-hydroxypropyl]piperaz-ine, N 2 —[N 2 ,N 5 -Bis(aminopropyl)-L-ornithyl]-N—N-dioleyl-L-α-asparagine, N—[N 2 —[N 2 ,N 5 -Bis[(1,1-dimethylethoxy)carbonyl]-N 2 ,N 5 -bis[3-[(1,1-dimethylethoxy)carbonyl]-aminopropyl]-L-ornithyl-N—N-dioleyl-L-glutaminyl]-L-glutamic acid, 1,4-bis[(3-(3-aminopropyl)-oleylamino)propyl]piperazine, N 2 —[N 2 ,N 5 -Bis(3-aminopropyl)-L-omithyl]-N,N-dipalmityl-L-glutamine,N 2 —[N 2 ,N 5 -Bis(aminopropyl)-L-ornithyl]-N—N-dipalmityl-L-α-glutamine, N 2 —[N 2 ,N 5 -Bis(aminopropyl)-L-ornithyl]-N—N-dipalmityl-L-α-asparagine, N— [N 2 -[N 2 ,N 5 -Bis[(1,1-dimethylethoxy)carbonyl]-N 2 ,N 5 -bis[3-[(1,1-dimethylethoxy)-carbonyl]aminopropyl]-L-ornithyl-N—N-dipalmityl-L-glutaminyl]-L-glutamic acid, N 2 —[N 2 ,N 5 -Bis(3-aminopropyl)-L-ornithyl]-N,N-dimyristyl-L-glutamine, N 2 —[N 2 ,N 5 -Bis-(aminopropyl)-L-omithyl]-N—N-dimyristyl-L-α-glutamine, N 2 —[N 2 ,N 5 -Bis(aminopropyl)-L-omithyl]-N—N-dimyristyl-L-α-asparagine, 1,4-bis[(3-(3-amino-2-hydroxypropyl)-palmitylamino)-2-hydroxypropyl]piperazine, N—[N 2 —[N 2 ,N 5 -Bis[(1,1-dimethylethoxy)-carbonyl]-N 2 ,N 5 -bis[3-[(1,1-dimethylethoxy)carbonyl]aminopropyl]-L-ornithyl-N—N-dimyristyl-L-glutaminyl]-L-glutamic acid, 1,4-bis[(3-(3-aminopropyl)-myristylamino)-propyl]piperazine, N 2 —[N 2 ,N 5 -Bis(3-aminopropyl)-L-omithyl]-N,N-dilaureyl-L-glutamine, N 2 —[N 2 ,N-Bis(aminopropyl)-L-ornithyl]-N—N-dilaureyl-L-α-glutamine, N 2 —[N 2 ,N-Bis(aminopropyl)-L-ornithyl]-N—N-dilaureyl-L-α-asparagine, N—[N 2 —[N 2 ,N 5 -Bis[(1,1-dimethylethoxy)carbonyl]-N 2 ,N 5 -bis[3-[(1,1-dimethylethoxy)carbonyl]amino-propyl]-L-omithyl-N—N-dilaureyl-L-glutaminyl]-L-glutamic acid, 3-[N′,N″-bis(2-tert-butyloxycarbonylaminoethyl)guanidino]-N,N-dioctadec-9-enylpropionamide, 3-[N′,N″-bis(2-tertbutyloxycarbonylaminoethyl)guanidino]-N,N-dipalmitylpropionamide, 3-[N′,N″-bis(2-tertbutyloxycarbonylaminoethyl)guanidino]-N,N-dimyri stylpropionamide, 1,4-bis[(3-(3-aminopropyl)-palmitylamino)propyl]piperazine, 1,4-bis[(3-(3-amino-2-hydroxypropyl)-oleylamino)propyl]piperazine, N,N-(2-hydroxy-3-aminopropyl)-N-2-hydroxypropyl-3-N,N-diolylaminopropane, N,N-(2-hydroxy-3-aminopropyl)-N-2-hydroxypropyl-3-N,N-dipalmitylaminopropane, N,N-(2-hydroxy-3-aminopropyl)-N-2-hydroxypropyl-3-N,N-dimyri stylaminopropane, 1,4-bis[(3-(3-amino-2-hydoxypropyl)-myristylamino)propyl]piperazine, [(3-aminopropyl)-bis-(2-tetradecyloxyethyl)]methyl ammonium bromide, [(3-aminopropyl)-bis-(2-oleyloxyethyl)]methyl ammonium bromide, [(3-aminopropyl)-bis-(2-palmityloxyethyl)]methyl ammonium bromide, Oleoyl-2-hydroxy-3-N,N-dimethyamino propane, 2-didecanoyl-1-N,N-dimethylamino-propane, palmitoyl-2-hydroxy-3-N,N-dimethyamino propane, 1,2-dipalmitoyl-1-N,N-dimethylaminopropane, myristoyl-2-hydroxy-3-N,N-dimethyamino propane, 1,2-dimyristoyl-1-N,N-dimethylaminopropane, (3-Amino-propyl)-&gt;4-(3-amino-propyl-amino)-4-tetradecylcarbamoyl-butylcarbamic acid cholesteryl ester, (3-Amino-propyl)-&gt;4-(3-amino-propylamino-4-carbamoylbutylcarbamic acid cholesteryl ester, (3-Amino-propyl)-&gt;4-(3-amino-propylamino)-4-(2-dimethylamino-ethylcarbamoyl)-butylcarbamic acid cholesteryl ester, Spermine-5-carboxyglycine (N′-stearyl-N′-oleyl) amide tetratrifluoroacetic acid salt, Spermine-5-carboxyglycine (N′-stearyl-N′-elaidyl) amide tetratrifluoroacetic acid salt, Agmatinyl carboxycholesterol acetic acid salt, Spermine-5-carboxy-(3-alanine cholesteryl ester tetratrifluoroacetic acid salt, 2,6-Diaminohexanoeyl 3-alanine cholesteryl ester bistrifluoroacetic acid salt, 2,4-Diaminobutyroyl β-alanine cholesteryl ester bistrifluoroacetic acid salt, N,N-Bis (3-aminopropyl)-3-aminopropionyl β-alanine cholesteryl ester tristrifluoroacetic acid salt, [N,N-Bis(2-hydroxyethyl)-2-aminoethyl]aminocarboxy cholesteryl ester, Stearyl carnitine ester, Palmityl carnitine ester, Myristyl carnitine ester, Stearyl stearoyl carnitine ester chloride salt, L-Stearyl Stearoyl Carnitine Ester, Stearyl oleoyl carnitine ester chloride, Palmityl palmitoyl carnitine ester chloride, Myristyl myristoyl carnitine ester chloride, L-Myristyl myristoyl carnitine ester chloride, 1,4-bis[(3-(3-amino-2-hydroxypropyl)-palmitylamino)-propyl]piperazine, N-(3-aminopropyl)-N,N′-bis-(dodecyloxyethyl)-piperazinium bromide, N-(3-aminopropyl)-N,N′-bis-(oleyloxyethyl)-piperazinium bromide, N-(3-aminopropyl)-N,N′-bis-(palmityloxyethyl)-piperazinium bromide, N-(3-aminopropyl)-N,N′-bis-(myristyloxyethyl)-piperazinium bromide, N-(3-aminopropyl)-N′-methyl-N,N′-(bis-2-dodecyloxyethyl)-piperazinium bromide, N-(3-aminopropyl)-N′-methyl-N,N′-(bis-2-oleyloxyethyl)-piperazinium bromide, N-(3-aminopropyl)-N′-methyl-N,N′-(bis-2-palmityloxyethyl)-piperazinium bromide, N-(3-aminopropyl)-N′-methyl-N,N′-(bis-2-myristyloxyethyl)-piperazinium bromide, 1,4-bis[(3-(3-aminopropyl)-oleylamino)-2-hydroxy-propyl]piperazine, 1,4-bis[(3-(3-aminopropyl)-myristylamino)-2-hydroxy-propyl]piperazine, or 1,4-bis[(3-(3-aminopropyl)-palmitylamino)-2-hydroxy-propyl]piperazine, 3-alkyloxy-2-hydroxy-1-histidylamidopropane, 3-alkyloxy-2-hydroxy-1-aminopropane, 4-alkyloxy-3-hydroxy-1-histidylamidopropane, 4-alkyloxy-3-hydroxy-1-aminopropane, 5-alkyloxy-4-hydroxy-1-histidylamidopropane, 5-alkyloxy-4-hydroxy-1-aminopropane, 6-alkyloxy-5-hydroxy-1-hi stidylamidopropane, 6-alkyloxy-4-hydroxy-1-aminopropane; 2,3-dialkoxy-1,4-bis(N-methyl-N-carboxyspermineamido)-aminobutane, 2,3-dialkoxy-1,4-bis(N-methyl-N-hi stidinylamido)aminobutane, 2,3-dialkoxy-1,4-bis(N-methyl-N-arginylamido)aminobutane, 2,3-dialkoxy-1,4-bis(N-methyl-N-lysinylamido)aminobutane, 2,3-dialkoxy-1,4-bis(N-methyl-N′-ornithinyl-amido)aminobutane, 2,3-dialkoxy-1,4-bis(N-methyl-N-serinylamido)aminobutane, 2,3-dialkoxy-1,4-bis(N-methyl-N-homoerinylamido)aminobutane, 2,3-dialkoxy-1,4-bis(N-methyl-N-(diaminobutanyl)amido)aminobutane, 2,3-dialkoxy-1,4-bis(N-methyl-N-(di-aminopropyl)amido)aminobutane, 2,3-dialkoxy-1,4-bis(N-methyl-N-(2-hydroxylpropyl-amine))aminobutane, 2,3-dialkoxy-1,4-bis(N-methyl-N-(2-diaminopropyl))aminobutane, 2,3-dialkoxy-1,4- and bis(N-methyl-N-propylamine)aminobutane. The alkoxy in the above list may be myristyloxy, myristeleyloxy lauryloxy, palmityloxy, palmitoleyloxy, oleyloxy and streaylox. 
     In some embodiments transfection agents contains lyso-phosphatidylcholine, sphingomyelin, disaturated phosphatidylcholine, saturated and unsaturated phosphatidylcholine, disaturated phosphatidylethanolamine, phosphatidylethanolamine saturated and unsaturated, phosphatidylserine, phosphatidylinositol, lyso-bis phosphatidic acid, cholesterol, and diglyceride. Helper Lipids may include, complete exosomes solutions, total extract of lipids isolated from exosomes. The Helper lipids can be formulated into exosome like lipid particles so that lyso-phosphatidylcholine is from 3.75 to 6.21% of total lipid content, a sphingomyelin, a sphingosin, or ceramide is from 8.26 to 12, 41% of total, phosphatidylcholine-disaturated is from 3.00 to 4.81% of total, Phosphatidylethanolamine-mix is from 12.00 to 19.22%, phosphatidylserine is from 5.17 to 6.89%, phosphatidylinositol is from 5.17 to 6.89%, Phosphatidylethanolamine disaturated is from 2.61 to 2.85%, Phosphatidylethanolamine mix is from 13.42 to 14.65%, cholesterol is from 13.01 to 16.61%, diglycerides are from 4.76 to 7.05% and cationic lipids can be the remaining % in such formulations. Cationic lipids can be substituted or used in combination with a sphingomyelin, a sphingosin, or ceramides. Depending on cell type the % composition of helper lipids and can be adjusted to optimize formulations of synthetic exosomes with various transfection enhancers. 
     When the composition contains a neutral lipid, that lipid a saturated or unsaturated, or mixed acyl phospatidyl ethanol mine (PE) or phospahtidyl choline (PC), for example, DOPE, DPhPE, saturated and unsaturated DPPE, saturated and unsaturated DMPE, cholesterol, DOPC, Lyso-PE (1-acyl-2-hydroxy-sn-glycero-3-phosphoethanolamine), Lyso-PC (1-acyl-3-hydroxy-sn-glycero-3-phosphocholine), 3-alkyloxy-2-hydroxy-1-acetamidopropane, 4-alkyloxy-3-hydroxy-1-acetamidopropane, 5-alkyloxy-4-hydroxy-1-acetamidopropane or 6-alkyloxy-5-hydroxy-1-acetamidopropane. The alkyloxy in the above list may be myristyloxy, myristeleyloxy lauryloxy, palmityloxy, palmitoleyloxy, oleyloxy or streayloxy. In some embodiments, the tranfection agent. may contain more than one of these neutral lipids or exosomes, lipids from exosomes or total lipid extracts from exosomes compositions. 
     In other embodiments the transfection agent comprises at least one polyamine moiety. Suitable polyamines include dense star dendrimers, PAMAM dendrimers, NH 3  core dendrimers, ethylenediamine core dendrimers, dendrimers of generation 5 or higher, dendrimers with substituted groups, dendrimers having one or more amino acids, grafted dendrimers, activated dendrimers, polyethylenimine, and polyethylenimine conjugates, polycationic peptides such as polylysine, polyorinthine, polyhistidine, polyarginine 
     In other embodiments, cell surface ligand containing adhesion peptide sequences is conjugated to a nucleic acid binding group. In some of these embodiments, the nucleic acid binding group is linked to a polyamine or peptide nucleic acid. The polyamine optionally comprises at least one spermine moiety. 
     Suitable cell surface ligands containing adhesion peptide sequences that are derived from cell adhesion proteins include, but are not limited to, a sequence selected from the group consisting of SEQ ID NOs:202-503, as set forth in Table 1, below, or in the sequence listings. 
     In some embodiments, the peptides of SEQ ID NOs:202-503 are optionally linked to a nucleic acid binding moiety, a helper lipid, a cationic lipid, a cationic polymer, a GPI anchor peptide or other chemical moieties that associate with transfection complexes. 
     In some embodiments, the peptides of SEQ ID NOs:202-500 are used with other surface ligands, such as antibodies, antibody fragments, single chain antibodies, aptemers, or peptides from phage display. In certain embodiments, these surface ligands are optionally attached to nucleic acid binding moieties, to lipids or lipid associating moieties. 
     In particular embodiments, the transfection agent comprises at least one cationic lipid, and optionally also contains in various combinations with one or more neutral and/or helper lipids, targeting moieties, cell penetration agent, fusion agents, and lysomotropic agents. 
     In some embodiments, the presently disclosed complexes comprise one more agents selected from the group consisting of fusogenic agents, nuclear localization sequences, cell penetration agent, transport peptides, receptor-ligand or cell adhesion peptides. 
     It is to be understood that while some peptides are disclosed herein in the context of one particular use, all of the peptides presently disclosed can be used in other uses as well. Thus, by way of example only, the peptides of SEQ ID NOs:1-41 are disclosed to be nuclear localization peptides. However, these peptides can be used as a nucleic acid binding peptide, a nucleic acid condensing peptide, a transfection enhancer. 
     In specific embodiments, the cell surface ligand containing adhesion peptide sequences is covalently linked to the transfection agents, the cationic lipid, the neutral lipid, helper lipid, a chemical group that associates with lipids or liposomes, and/or the polyamine. 
     In specific embodiments, the plant virus movement protein or peptide derived from plant virus movement proteins is covalently linked to the transfection agents, the cationic lipid, the neutral lipid, helper lipid, a chemical group that associates with lipids or liposomes, and/or the polyamine. 
     In other embodiments, the plant virus movement protein or peptide derived from plant virus movement proteins is conjugated to a nucleic acid binding group. In some of these embodiments, the nucleic acid binding group is linked to a polyamine or peptide nucleic acid. The polyamine optionally comprises at least one spermine moiety. 
     Suitable virus movement proteins and peptides derived from plant virus movement proteins sequences include, but are not limited to, a sequence selected from the group consisting of SEQ ID NOs:506-580, as set forth in Table 1, below, or in the sequence listings. 
     In some embodiments, the peptides of SEQ ID NOs:506-580 are optionally linked to a nucleic acid binding moiety, a helper lipid, a cationic lipid, a cationic polymer, a GPI anchor peptide or other chemical moieties that associate with transfection complexes. 
     In some embodiments, the peptides of SEQ ID NOs:506-580 are used with other surface ligands, such as antibodies, antibody fragments, single chain antibodies, aptamers, or peptides from phage display. In certain embodiments, these surface ligands are optionally attached to nucleic acid binding moieties, to lipids or lipid associating moieties. 
     In particular embodiments, the transfection agent comprises at least one cationic lipid, and optionally also contains in various combinations with one or more neutral and/or helper lipids, targeting moieties, surface ligand, fusion agents, and lysomotropic agents. 
     In some embodiments, the presently disclosed complexes comprise one more agents selected from the group consisting of fusogenic agents, cell penetration agents, nuclear localization sequences, transport peptides, plant movement protein or peptide derived from, receptor-ligand, surface ligand or cell adhesion peptides. 
     It is to be understood that while some peptides are disclosed herein in the context of one particular use, all of the peptides presently disclosed can be used in other uses as well. Thus, by way of example only, the peptides of SEQ ID NOs:1-41 are disclosed to be nuclear localization peptides. However, these peptides can be used as a nucleic acid binding peptide, a nucleic acid condensing peptide, a transfection enhancer. 
     In another aspect, disclosed herein are pharmaceutical compositions containing a complex as described herein, and a pharmaceutically acceptable carrier. 
     In another aspect, disclosed herein are methods of transfecting a cell, by contacting a cell with a complex as described herein. In some embodiments, the cell is selected from the group consisting of a primary cell culture, a passaged cell culture, suspension cell line and an attached cell line. Suitable cells include all human cell lines and all animal cell lines. In some embodiments, the cell is a blood derived cell or the cell is a stem cell, while in other embodiments, the cell is a neuron. 
     In one method, a nucleic acid, protein or, peptide, or pharmaceutical is contacted with a cell surface ligand containing adhesion peptide, the plant virus movement protein, or peptide derived therefrom, capable associating with nucleic acid, protein, peptide or pharmaceutical and the resulting mixture is added to a transfection agent then contacted to cells. 
     In one embodiment of the transfection methods, cell surface ligand containing adhesion peptide, the plant virus movement protein, or peptide derived therefrom, are contacted with a transfection agent capable of associating with a nucleic acid, a protein, a peptide or a pharmaceutical composition, followed by addition of a nucleic acid, a protein, peptide or pharmaceutical then contacted to cells. 
     In another method, cell surface ligand containing adhesion peptide, the plant virus movement protein, or peptide derived therefrom, linked to a nucleic acid binding moiety is contacted with transfection agent capable of associating with nucleic acid, protein, peptide or pharmaceutical followed by addition of a nucleic acid, protein, peptide or pharmaceutical then contacted to cells 
     In another method, cell surface ligand containing adhesion peptide, the plant virus movement protein, or peptide derived therefrom, contacted with transfection agent followed by addition of a fusion agent and then contacted with a nucleic acid, protein, peptide or pharmaceutical then contacted to cells 
     In another method, cell surface ligand containing adhesion peptide, the plant virus movement protein, or peptide derived therefrom, linked to a nucleic acid binding moiety is contacted with transfection agent followed by addition of a fusion agent and then contacted with a nucleic acid, protein, peptide or pharmaceutical then contacted to cells. 
     In another method cell surface ligand containing adhesion peptide, the plant virus movement protein, or peptide derived therefrom, is contacted with a fusion agent followed by addition of a transfection agent and then contacted with a nucleic acid, protein, peptide or pharmaceutical then contacted to cells. 
     In another method cell surface ligand containing adhesion peptide, the plant virus movement protein, or peptide derived therefrom, linked to a nucleic acid binding moiety is contacted with a fusion agent followed by addition of a transfection agent and then contacted with a nucleic acid, protein, peptide or pharmaceutical then contacted to cells. 
     In another method cell surface ligand containing adhesion peptide, the plant virus movement protein, or peptide derived therefrom, linked to a nucleic acid binding moiety is contacted with a transfection agent, then contacted with a nucleic acid, protein, peptide or pharmaceutical, then contacted with a fusion agent, and then contacted to cells. 
     In another method cell surface ligand containing adhesion peptide, the plant virus movement protein, or peptide derived therefrom, linked to a nucleic acid binding moiety is contacted with a transfection agent containing exosomes, or lipid extracts from exosomes, then contacted with a nucleic acid, protein, peptide or pharmaceutical, then contacted with a fusion agent, and then contacted to cells 
     In another method an aqueous solution of transfection enhancers, that may optionally contain one or more a plant virus movement proteins or peptides derived from plant virus movement proteins linked to a nucleic acid binding moiety, that may optionally contain buffers, that may optionally contain one or more cell surface ligands, that may optionally contain one or more nuclear localization agents, that may optionally contain one or more cell penetration peptides, that may optionally containing one or more fusion peptides or proteins, all or some of these transfection enhancers may optionally be linked to nucleic acid binding domain is mixed with an ethanol or aqueous solution of lipids that may contain cationic lipids, helper lipids, exosomes or total lipid extracts of exosomes or various mixtures of such lipids or lipid particles The mixture is then contacted with nucleic acids, protein, peptides, exosomes, growth enhancers, and then cells. 
     In another method an ethanol solution of transfection enhancers, that may optionally contain one or more a plant virus movement proteins or peptides derived from plant virus movement proteins linked to a nucleic acid binding moiety, that may optionally contain buffers, that may optionally contain one or more cell surface ligands, that may optionally contain one or more nuclear localization agents, that may optionally contain one or more cell penetration peptides, that may optionally containing one or more fusion peptides or proteins, all or some of these transfection enhancers may optionally be linked to nucleic acid binding domain is mixed with an ethanol or aqueous solution of lipids that may contain cationic lipids, helper lipids, exosomes or total lipid extracts of exosomes or various mixtures of such lipids or lipid particles The mixture is then contacted with nucleic acids, protein, peptides, exosomes, growth enhancers, and then cells. 
     In another method an aqueous solution of transfection enhancers, that may optionally contain one or more a plant virus movement proteins or peptides derived from plant virus movement proteins linked to a nucleic acid binding moiety, that may optionally contain buffers, that may optionally containing one or more cell surface ligands, that may optionally one or more nuclear localization agents, that may optionally contain one or more cell penetration peptides, all or some of these transfection enhancers may optionally be linked to nucleic acid binding domain is mixed with an ethanol or aqueous solution of lipids that may contain cationic lipids, helper lipids, exosomes or total lipid extracts of exosomes or various mixtures of such lipids or lipid particles. This mixture is then contacted with one or more fusion peptides or proteins which may optionally be in a buffer. The mixture is then contacted with nucleic acids, protein, peptides, exosomes, growth enhancers, and then cells. 
     In another method, a transfection complex is contacted with a nucleic acid, protein or, peptide then contacted with cell surface ligand containing adhesion peptide, the plant virus movement protein, or peptide derived therefrom, and then contacted to cells. 
     Those skilled in the art understand that concentrations of the various components, such as cationic lipid, helper lipid, the plant virus protein or peptide derived therefrom, cell surface ligand, fusogenic reagent, nuclear ligand, cationic polymer, condensing agent, cell penetration, lysomotrophic agent, exosomes or exosome lipids or proteins from exosomes, peptides derived for exosomal proteins and transfection enhancers are optimized according to the cell to be transfected or the in vivo application. In some embodiments, the concentration of each component ranges from 0.001 to 20 mg/mL, depending on solubility and formulation solvent. 
     In some embodiments, the plant virus movement protein derived peptides disclosed herein are suitable for use in the disclosed transfection enhancers to enhance transfection into HeLa cells, HuVec cells, iPS cells, NL-1 cells, C2C12 cells, human fibroblast cells, stem cells, Jurkat cells, rat cortical neurons, THP-1 cells, and human skeletal muscle cells, among others. 
     In some embodiments, the peptide useful to enhance the transfection of HeLa cells is selected from the group consisting of SEQ ID NOs: 107, 205, 216, 218, 219, 220, 224, 226, 229, 230, 234, 236, 236, 237, 238, 239, 256, 268, 323, 326, 327, 328, 332, 335, 336, 338, 341, 342, 343, 344, 345, 347, 348, 349, 350, 351, 352, 353, 354, 355, 357, 358, 359, 360, 361, 363, 365, 367, 369, 371, 373, 375, 377, 379, 381, 383, 450, 452, 454, and 503 can be used in combination with the plant virus movement protein or peptide derived therefrom. In some of these embodiments, the peptide useful to enhance the transfection of HeLa cells is selected from the group consisting of SEQ ID NOs:205, 237, 268, 326, 328, 335, 336, 342, 347, 348, 352, 353, 355, 357, 358, 365, 367, 377, 379, 381, 450, 454, and 503 can be used in combination with the plant virus movement protein or peptide derived therefrom. In some embodiments, the peptide useful to enhance the transfection of HuVec cells is selected from the group consisting of SEQ ID NOs:205, 216, 218, 219, 220, 224, 226, 229, 230, 234, 236, 236, 237, 238, 239, 256, 268, 323, 326, 327, 328, 332, 335 336, 338, 341, 342, 343, 344, 345, 347, 348, 349, 350, 351, 352, 353, 354, 355, 357, 358, 359, 360, 361, 363, 365, 367, 369, 371, 373, 375, 377, 379, 381, 383, 450, 452, 454, and 503 can be used in combination with the cell surface ligand containing adhesion peptide, the plant virus movement protein, or peptide derived therefrom. In some of these embodiments, the peptide useful to enhance the transfection of HuVec cells is selected from the group consisting of SEQ ID NOs:236, 358, 373 can be used in combination with the cell surface ligand containing adhesion peptide, the plant virus movement protein, or peptide derived therefrom. 
     In some embodiments, the peptide useful to enhance the transfection of NL-1 iPS cells is selected from the group consisting of SEQ ID NOs: 107, 205, 216, 218, 219, 220, 224, 226, 229, 230, 234, 236, 236, 237, 238, 239, 256, 268, 323, 326, 327, 328, 332, 335, 336, 338, 341, 342, 343, 344, 345, 347, 348, 349, 350, 351, 352, 353, 354, 355, 357, 358, 359, 360, 361, 363, 365, 367, 369, 371, 373, 375, 377, 379, 381, 383, 450, 452, 454, 503 can be used in combination with the cell surface ligand containing adhesion peptide, the plant virus movement protein, or peptide derived therefrom. In some of these embodiments, the peptide useful to enhance the transfection of NL-1 iPS cells is selected from the group consisting of SEQ ID NOs:216, 224, 226, 236, 236, 323, 327, 341, 343, 347, 348, 349, 350, 351, 354, 358, 360, 373, 383, 450, 454, 503 can be used in combination with the cell surface ligand containing adhesion peptide, the plant virus movement protein, or peptide derived therefrom. 
     In some embodiments, the peptide useful to enhance the transfection of C2C12 cells is selected from the group consisting of SEQ ID NOs:205, 216, 218, 219, 220, 224, 226, 229, 230, 234, 236, 236, 237, 238, 239, 256, 268, 323, 326, 327, 328, 332, 335, 336, 338, 341, 342, 343, 344, 345, 347, 348, 350, 351, 352, 353, 354, 355, 357, 358, 359, 360, 361, 363, 365, 367, 369, 371, 373, 375, 377, 379, 381, 383, 450, 452, 454, 501, 502, 503 can be used in combination with the cell surface ligand containing adhesion peptide, the plant virus movement protein, or peptide derived therefrom. In some of these embodiments, the peptide useful to enhance the transfection of C2C12 cells is selected from the group consisting of SEQ ID NOs:218, 230, 237, 239, 256, 323, 326, 328, 335, 336, 342, 343, 345, 347, 348, 352, 357, 359, 367, 375, 379, 381, 450, 452, 454, 501, 502, 503 can be used in combination with the cell surface ligand containing adhesion peptide, the plant virus movement protein, or peptide derived therefrom, 
     In some embodiments, the peptide useful to enhance the transfection of human fibroblast cells is selected from the group consisting of SEQ ID NOs:205, 216, 218, 219, 220, 224, 226, 229, 230, 234, 236, 236, 237, 238, 239, 256, 268, 323, 326, 327, 328, 332, 335, 336, 338, 341, 342, 343, 344, 345, 347, 348, 349, 350, 351, 352, 353, 354, 355, 357, 358, 359, 360, 361, 363, 365, 367, 369, 371, 373, 375, 377, 379, 381, 383, 450, 452, 454, 501, 503 can be used in combination with cell surface ligand containing adhesion peptide, the plant virus movement protein, or peptide derived therefrom. In some of these embodiments, the peptide useful to enhance the transfection of human fibroblast cells is selected from the group consisting of SEQ ID NOs:205, 218, 219, 229, 230, 335, 336, 342, 344, 348, 349, 350, 351, 353, 355, 357, 361, 367, 369, 375, 379, 381, 450, 454, 501, 503 can be used in combination with the cell surface ligand containing adhesion peptide, the plant virus movement protein, or peptide derived therefrom. 
     In some embodiments, the peptide useful to enhance the transfection of Jurkat cells is selected from the group consisting of SEQ ID NOs:205, 216, 218, 219, 220, 224, 226, 229, 230, 234, 236, 236, 237, 238, 239, 256, 268, 323, 326, 327, 328, 332, 335, 336, 338, 341, 342, 343, 344, 345, 347, 348, 349, 350, 351, 352, 353, 354, 355, 357, 358, 359, 360, 361, 363, 365, 367, 369, 371, 373, 375, 377, 379, 381, 383, 450, 452, 454, 501, 503 can be used in combination with the cell surface ligand containing adhesion peptide, the plant virus movement protein, or peptide derived therefrom. In some of these embodiments, the peptide useful to enhance the transfection of Jurkat cells is selected from the group consisting of SEQ ID NOs:218, 349, 358 can be used in combination with the cell surface ligand containing adhesion peptide, the plant virus movement protein, or peptide derived therefrom. 
     In some embodiments, the peptide useful to enhance the transfection of rat cortical neuron cells is selected from the group consisting of SEQ ID NOs: 220, 236, 238, 323, 327, 336, 338, 341, 343, 347, 348, 350, 351, 352, 354, 367, 369, 373, 375, 377, 454 can be used in combination with the cell surface ligand containing adhesion peptide, the plant virus movement protein, or peptide derived therefrom. 
     In some embodiments, the peptide useful to enhance the transfection of THP-1 cells is selected from the group consisting of SEQ ID NOs:219, 229, 230, 239, 323, 328, 332, 341, 343, 350, 351, 357, 358, 375, 450, 454 can be used in combination with the cell surface ligand containing adhesion peptide, the plant virus movement protein, or peptide derived therefrom. 
     In some embodiments, the peptide useful to enhance the transfection of human skeletal muscle cells is selected from the group consisting of SEQ ID NOs: 218, 219, 230, 328, 336, 344, 350, 351, 353, 355, 365, 375 can be used in combination with the cell surface ligand containing adhesion peptide, the plant virus movement protein, or peptide derived therefrom. 
     In some embodiments combination of different surface ligands that have preference for a given cell types are mixed together to target multiple cell types in a single formulation. 
     In another aspect, disclosed herein are kits containing a transfection agent and a peptide or protein or a modified peptide or modified protein with cell surface ligand containing adhesion peptide, the plant virus movement protein, or peptide derived therefrom, as described herein. In some embodiments, the kit further comprises instructions for the preparation and the use of the transfection complexes. In certain embodiments, the kit further comprises separate compartments for the various components. In certain embodiments transfection agents may comprise various peptides selected from SEQ ID NOs 1-505 in combination with SEQ ID NOs 506-580 that are suitable for transfecting cells in vivo or in vitro. 
     EXAMPLES 
     Example 1: Surface Ligands 
     Cells were plated to so that on the day of transfection the cells were 70% confluent in 96 well tissue culture plates. A DOMTA/DOPE Lipid solution (1:1 molar ratio) at 2 mg/mL in water were mixed with an equal volume cell surface ligand containing adhesion peptide, the plant virus movement protein, or peptide derived therefrom, that had the nucleic acid binding moiety RRRRRRRRRRRR (SEQ ID NO: 109) or KKKKKKKKKKKKKKKK (SEQ ID NO: 102) covalently linked to the N-terminus, or C-terminus of each peptide during peptide synthesis. Peptides were at 2 mg/mL in water. The DOTMA/DOPE and the solution containing the cell surface ligand containing adhesion peptide, the plant virus movement protein, or peptide derived therefrom, surface ligand peptide the was diluted 1 to 1 (v/v) in water. DOMTA/DOPE/PEPTIDE solutions were added to 0.1 mL of a plasmid DNA solution (EF 1Alpha eGFP plasmid) at 10 μg/mL in OptiMEM. 
     All solutions were at room temperature. A volume of 0.2 mL of a 10 μg/mL solution of DNA was aliquoted into each well of a non-tissue cultured treated plate. 1-12 μL of transfection reagents were added to DNA solutions respectively. The transfection reagent and DNA solution was mixed by pipetting up and down twice. Transfection complexes were formed for 10 minutes. After 10 minutes, 0.01 or 0.02 mL of the transfection complex was added to cells. HuVEC, HeLa, human adult keratinocytes, human primary adult fibroblast, and A549 cells all respectively received 0.01 mL of the transfection complex. Human skeletal muscle cells, mouse C2C12 cells, and Jurkat, cells received 0.02 mL of the transfection complex. Cells were incubated for 42 hours at 37° C. at 5% CO 2 . Plates were read on a fluorescent plate reader. Cells were also examined visually under a microscope to assess the extent of transfection (in terms of the percent of cells transfected) with a flouresenct microscope. Other modes of analysis, for example quantification with B-galactocidase or luciferase reports plasmids can also be used. If the plate reader did not show a sufficient signal to noise ratio, then plates were scored for cells transfected and those peptides that show increase over DOTMA alone were noted. 
     Not all surface ligands increased transfection efficiency even though they were suggested to be used to attach cell to tissue culture plates. Surface ligand that caused increase expression of GFP over the lipid DOTMA with no surface ligand were considered to enhance transfection or had higher % cells transfected were considered to enhance transfection. Peptides that gave greater than 2-fold enhancement are noted in bold and could be further optimized by adding different transfection enhancers or could be used with different cationic lipids or polymers. 
     Tables 2-7, below, provide the results of the transfection enhancements with the various cell types, while Table 8 lists the peptides that were determined by visual inspection to enhance the transfection of the denoted cell lines over the lipid DOTMA. 
     The following sequences increased transfection expression and the % cells transfected over the no peptide control on some examined cell types: SEQ ID NOs: 205, 216, 218, 219, 220, 226, 229,230, 236, 237, 238, 239, 256, 268, 323, 326, 327, 328, 332, 336, 338, 341, 342, 343, 344, 345, 347, 348, 349, 350, 351, 352, 353, 354, 355, 357, 358, 359, 360, 361, 365, 367, 369, 373, 375, 377, 379, 381, 393, 450, 452, 454, 501, and 503. 
     Various DOTMA/peptide transfection reagents that showed enhanced transfection efficiency over DOTMA alone were further formulated with the fusogenic peptide (SEQ ID NO:72). The fusogenic peptide was added to the DOMTA/peptide formulation to achieve a final concentration of 0.1 mg/mL to see if transfection reagents were further enhanced by the addition of a fusogenic peptide and could provide higher transfection efficiency into HeLa cells and expression than the commercially available Lipofectamine 2000. The results are shown in Table 9. 
     The following Table 1 lists the peptide sequences that are referenced herein. 
     
       
         
           
               
               
             
               
                 TABLE 1 
               
               
                   
               
               
                 SEQ 
                   
               
               
                 ID 
                   
               
               
                 NO 
                 Sequence 
               
               
                   
               
             
            
               
                   
               
            
           
           
               
               
            
               
                 1 
                 GYSTPPKKKRKVEDP 
               
               
                   
               
               
                 2 
                 GYSTPPKTRRRP 
               
               
                   
               
               
                 3 
                 GYSTPGRKKR 
               
               
                   
               
               
                 4 
                 GYSTPRRNRRRRW 
               
               
                   
               
               
                 5 
                 PDEVKRKKKPPTSYG 
               
               
                   
               
               
                 6 
                 PRRRTKPPTSYG 
               
               
                   
               
               
                 7 
                 RKKRGPTSYG 
               
               
                   
               
               
                 8 
                 WRRRRNRRPTSYG 
               
               
                   
               
               
                 9 
                 GYGPPKKKRKVEAPYKA 
               
               
                   
               
               
                 10 
                 PAAKRVKLD 
               
               
                   
               
               
                 11 
                 RQRRNELKRSP 
               
               
                   
               
               
                 12 
                 KRPAATKKAGQAKKKK 
               
               
                   
               
               
                 13 
                 VRKKRKTEEESPLKDKDAKKSKQE 
               
               
                   
               
               
                 14 
                 RLRRDAGGRGGVYEHLGGAPRRRK 
               
               
                   
               
               
                 15 
                 KRKGDEVDGVDECAKKSKK 
               
               
                   
               
               
                 16 
                 NQSSNFGPMKGGNFGGRSSGPYGGGGQYFAKPRNQGGY 
               
               
                   
               
               
                 17 
                 GGKRTADGSEFESPKKARKVEAYPKAW 
               
               
                   
               
               
                 18 
                 GGKRTADGSEFESPKKKRAVEAYPKAW 
               
               
                   
               
               
                 19 
                 GGKRTADGSEFESPKKKAKVEAYPKAW 
               
               
                   
               
               
                 20 
                 GGKRTADGSEFESPKKKRKVEAPYKAWK 
               
               
                   
               
               
                 21 
                 GGKRTADGSEFESPKKKRKVEYKAWK 
               
               
                   
               
               
                 22 
                 GYGPAAKRVKLDEAYPKAWK 
               
               
                   
               
               
                 23 
                 GGKRTADGSEFEPAAKRVKLDEAYPKAWK 
               
               
                   
               
               
                 24 
                 GTGPKKKRKVGGGGYGPKKKRLVG 
               
               
                   
               
               
                 25 
                 KRPAATKKAGQAKKKKLEAYPKAWK 
               
               
                   
               
               
                 26 
                 ATKGTKRSYEQMETGE 
               
               
                   
               
               
                 27 
                 GKWERKPIRCAS 
               
               
                   
               
               
                 28 
                 GYGKRTADSQHSTPPKKKRKVEAPYKAWK 
               
               
                   
               
               
                 29 
                 KRTADSQHSTPPKKKRKVEAPYKAWK 
               
               
                   
               
               
                 30 
                 GYGPPKKKRKVEAPYKAWKWAKYPAMRRAHHRRRRAS 
               
               
                   
                 HRRRTTTGT 
               
               
                   
               
               
                 31 
                 GYGPPKKKRKVEAPYKAWKRGARRYSKMKRRRRRVARRHRRRP 
               
               
                   
               
               
                 32 
                 FWGYGYGPPKKKRKVEAPYKAWK 
               
               
                   
               
               
                 33 
                 GKPSSDDEATADSQHSTPPKKKERKVED 
               
               
                   
               
               
                 34 
                 GKPTADDQHSTPPKKKRKVED 
               
               
                   
               
               
                 35 
                 GGKRTADGSEFESPKKARKVEAYPKAK 
               
               
                   
               
               
                 36 
                 EKIRLRPGRKKRYRLKHL 
               
               
                   
               
               
                 37 
                 PEGTRQARRNRRRRWRKR 
               
               
                   
               
               
                 38 
                 PEGTRQPRRNRRRRWRKR 
               
               
                   
               
               
                 39 
                 GVKRSYGAARGDDRRRPNVVAPYKAW 
               
               
                   
               
               
                 40 
                 KSVPNRTRTYIKLKRLRFKGAPYKAW 
               
               
                   
               
               
                 41 
                 EMRRRREEEGLQLRKQKREEQLFKRRN 
               
               
                   
               
               
                 42 
                 FEAALAEALAEALA 
               
               
                   
               
               
                 43 
                 Ac-LARLLPRLLARL-NHCH 3   
               
               
                   
               
               
                 44 
                 GLLEELLELLEELWEELLEG 
               
               
                   
               
               
                 45 
                 GWEGLIEGIEGGWEGLIEG 
               
               
                   
               
               
                 46 
                 GLFEALAEFIEGGWEGLIEG 
               
               
                   
               
               
                 47 
                 GLFEALLELLESLWELLLEA 
               
               
                   
               
               
                 48 
                 GGYCLEKWMIVASELKCFGNTA 
               
               
                   
               
               
                 49 
                 GGYCLTRWMLIEAELKCFGNTAV 
               
               
                   
               
               
                 50 
                 WEAALAEALAEALAEHLAEALAEALEALAA 
               
               
                   
               
               
                 51 
                 GLFGAIAGFIENGWEGMIDGWYG 
               
               
                   
               
               
                 52 
                 GIGAVLKVLTTGLPALISWIKRKRQQ 
               
               
                   
               
               
                 53 
                 GRKKRRQRRRPPQ 
               
               
                   
               
               
                 54 
                 RQIKIWFQNRRMKWKK 
               
               
                   
               
               
                 55 
                 GWTLNSAGYLLGKINLKALAALAKKIL 
               
               
                   
               
               
                 56 
                 WEAKLAKALAKALAKHLAKALAKALKACEA 
               
               
                   
               
               
                 57 
                 GLFKALLKLLKSLWKLLLKA 
               
               
                   
               
               
                 58 
                 GLFRALLRLLRSLWRLLLRA 
               
               
                   
               
               
                 59 
                 GLFEALLELLESLYELLLEA 
               
               
                   
               
               
                 60 
                 GLFEALEELWEA 
               
               
                   
               
               
                 61 
                 GLFLLEEWLE 
               
               
                   
               
               
                 62 
                 GLFLLEEWLEK 
               
               
                   
               
               
                 63 
                 GLFEALLELLESLWELLLEAK 
               
               
                   
               
               
                 64 
                 Suc-GLFKLLEEWLE 
               
               
                   
               
               
                 65 
                 Suc-GLFKLLEEWLEK 
               
               
                   
               
               
                 66 
                 GLFEAIAEFIEGGWEGLIEG 
               
               
                   
               
               
                 67 
                 GLFKAIAKFIKGGWKGLIKG 
               
               
                   
               
               
                 68 
                 IRFKKTKLIASIAMALC 
               
               
                   
               
               
                 69 
                 ALAGTIIAGASLTFQVLDKVlEELGKVSRK 
               
               
                   
               
               
                 70 
                 GLFEAIEGFIENGWEGMIDGWYG 
               
               
                   
               
               
                 71 
                 GYICRRARGDNPDDRCT 
               
               
                   
               
               
                 72 
                 GLFEAIAEFIEGGWEGLIEGCA 
               
               
                   
               
               
                 73 
                 GLFHAIAHFIHGGWHGLIHGWWYG 
               
               
                   
               
               
                 74 
                 RRRQRRKKRGGDIMGEWGNEIFGAIAGFLG 
               
               
                   
               
               
                 75 
                 GLFEAIADFIENGWEGMIDGGG 
               
               
                   
               
               
                 76 
                 ALAGTIIAGASLTFQVLDKVlEELGKVSRKK 
               
               
                   
               
               
                 77 
                 IRFKKTKLIASTAMA 
               
               
                   
               
               
                 78 
                 GLWHLLLHLWRRLLRLLR 
               
               
                   
               
               
                 79 
                 KKIMLLLMTLLLVSLPLAQEQ 
               
               
                   
               
               
                 80 
                 GLFEALLELLESLWELLLEAWYG 
               
               
                   
               
               
                 81 
                 RLLRLLLRLWRRLLRLLR 
               
               
                   
               
               
                 82 
                 LLELELLELELLLELELLELELLLEL 
               
               
                   
               
               
                 83 
                 GLFEALLELLESLWELLLEARRRRRRRR 
               
               
                   
               
               
                 84 
                 GLFEALLELLESLWELLLEARRRRRR 
               
               
                   
               
               
                 85 
                 GLFEALLELLESLWELLLEAKKKKKKKK 
               
               
                   
               
               
                 86 
                 GLFEALLELLESLWELLLEAKKKKKK 
               
               
                   
               
               
                 87 
                 GLFEALLELLESLWELLLEAKK 
               
               
                   
               
               
                 88 
                 GLFEALLELLESLWELLLEAKKKK 
               
               
                   
               
               
                 89 
                 GLFEALLELLESLWELLLEAEE 
               
               
                   
               
               
                 90 
                 GLFEALLELLESLWELLLEAEEEE 
               
               
                   
               
               
                 91 
                 GLFEALLELLESLWELLLEAEEEEEE 
               
               
                   
               
               
                 92 
                 GLFEALLELLESLWELLL 
               
               
                   
               
               
                 93 
                 PLSSIFSRIGDPRGARRYAKMKRRRRRVARRHRRRP 
               
               
                   
               
               
                 94 
                 GPFHYFQFLFPPV 
               
               
                   
               
               
                 95 
                 GSSSWWQRWWPPW 
               
               
                   
               
               
                 96 
                 RRRQRRKKR 
               
               
                   
               
               
                 97 
                 KKKK 
               
               
                   
               
               
                 98 
                 KKKKKK 
               
               
                   
               
               
                 99 
                 KKKKKKKK 
               
               
                   
               
               
                 100 
                 KKKKKKKKKK 
               
               
                   
               
               
                 101 
                 KKKKKKKKKKKK 
               
               
                   
               
               
                 102 
                 KKKKKKKKKKKKKKKK 
               
               
                   
               
               
                 103 
                 KKKKKKKKKKKKKKKKKKKK 
               
               
                   
               
               
                 104 
                 KKKKKKKKKKKKKKKKKKKKKKKK 
               
               
                   
               
               
                 105 
                 RRRR 
               
               
                   
               
               
                 106 
                 RRRRRR 
               
               
                   
               
               
                 107 
                 RRRRRRRR 
               
               
                   
               
               
                 108 
                 RRRRRRRRRR 
               
               
                   
               
               
                 109 
                 RRRRRRRRRRRR 
               
               
                   
               
               
                 110 
                 RRRRRRRRRRRRRRRR 
               
               
                   
               
               
                 111 
                 RRRRRRRRRRRRRRRRRRRR 
               
               
                   
               
               
                 112 
                 RRRRRRRRRRRRRRRRRRRRRRRR 
               
               
                   
               
               
                 113 
                 YKA 
               
               
                   
               
               
                 114 
                 KKKKKKKKWKGGGGACYGLPHLFCG 
               
               
                   
               
               
                 115 
                 YKAKKKKKKKKWK 
               
               
                   
               
               
                 116 
                 KTPKKAKKPKTPKKAKKP 
               
               
                   
               
               
                 117 
                 KKAKKPAATRKSSKNPKKPKTVKPKKVAK 
               
               
                   
               
               
                 118 
                 RGARRYSKMKRRRRRVARRHRRRP 
               
               
                   
               
               
                 119 
                 TRQARRNRRRRWRERQRGSGSG 
               
               
                   
               
               
                 120 
                 KRPRGRPKGSKKNWRRRKRRASRRSPRRR 
               
               
                   
               
               
                 121 
                 KRGRGRPRKQPPKEPSEVPTPKRPRGRPKGSKNK 
               
               
                   
               
               
                 122 
                 KEKYEKDIAAYRAKGKPAAKKGVVKAEKSKKKK 
               
               
                   
               
               
                 123 
                 YKAKKKKKKKKKKWK 
               
               
                   
               
               
                 124 
                 KKKKKKKGGC 
               
               
                   
               
               
                 125 
                 YRARRRRRRRRWR 
               
               
                   
               
               
                 126 
                 YRARRRRRRRRRRWR 
               
               
                   
               
               
                 127 
                 KGDPKKPRGKMSSYAFFVQTCREEHKKKHPDASVNFSEFSKK 
               
               
                   
               
               
                 128 
                 KKQLKKQLKKQLKQWK 
               
               
                   
               
               
                 129 
                 KKSPKKSPKKSPKKSK 
               
               
                   
               
               
                 130 
                 KLSKLEKKSKLEK 
               
               
                   
               
               
                 131 
                 KLSKLEKKLSKLEKKSKLEK 
               
               
                   
               
               
                 132 
                 KSLKKSLKKSLKKSK 
               
               
                   
               
               
                 133 
                 KIRRRGKNKVAARTCRQRRTDR 
               
               
                   
               
               
                 134 
                 KIRRRGKNKVAAQNCRKRKLET 
               
               
                   
               
               
                 135 
                 KRRIRREKNKMAAAKCRNRRRELT 
               
               
                   
               
               
                 136 
                 KDRSNLLERHTR 
               
               
                   
               
               
                 137 
                 KRPAATKKAGQAKKKL 
               
               
                   
               
               
                 138 
                 RRRRRREEEE 
               
               
                   
               
               
                 139 
                 RRRRRREEEEEE 
               
               
                   
               
               
                 140 
                 RRRRRREEEEEEEE 
               
               
                   
               
               
                 141 
                 RRRRRRRREEEE 
               
               
                   
               
               
                 142 
                 RRRRRRRREEEEEE 
               
               
                   
               
               
                 143 
                 RRRRRRRREEEEEEEE 
               
               
                   
               
               
                 144 
                 RRRRRRRRRRRREEEE 
               
               
                   
               
               
                 145 
                 RRRRRRRRRRRREEEEEE 
               
               
                   
               
               
                 146 
                 RRRRRRRRRRRREEEEEE 
               
               
                   
               
               
                 147 
                 KLSKLEKK 
               
               
                   
               
               
                 148 
                 SKLEK 
               
               
                   
               
               
                 149 
                 KLSKLEKKLSKLEKK 
               
               
                   
               
               
                 150 
                 PKKKRKVGGGRGDSP 
               
               
                   
               
               
                 151 
                 LPHKSMPCG 
               
               
                   
               
               
                 152 
                 GACLQHKSMPCG 
               
               
                   
               
               
                 153 
                 YGLPHLFCG 
               
               
                   
               
               
                 154 
                 SERSMNFCG 
               
               
                   
               
               
                 155 
                 DHYSLYEDLERGTDK 
               
               
                   
               
               
                 156 
                 ISLPRTSGAQRASTTR 
               
               
                   
               
               
                 157 
                 EKLQTKYGLPHKVEFCG 
               
               
                   
               
               
                 158 
                 TRISESQAKPGD 
               
               
                   
               
               
                 159 
                 LVFFDY 
               
               
                   
               
               
                 160 
                 WGGNGPTTFDCSGYTKYVFAK 
               
               
                   
               
               
                 161 
                 INIGTTGWGDHYSLY 
               
               
                   
               
               
                 162 
                 YDNIHG 
               
               
                   
               
               
                 163 
                 AGWGKFLVGFGRV 
               
               
                   
               
               
                 164 
                 SIGYPLP 
               
               
                   
               
               
                 165 
                 TTHWGFTL 
               
               
                   
               
               
                 166 
                 HLQIQPYPQISG 
               
               
                   
               
               
                 167 
                 KLNIVSVNG 
               
               
                   
               
               
                 168 
                 RGH 
               
               
                   
               
               
                 169 
                 DNRIRLQAKAA 
               
               
                   
               
               
                 170 
                 KIKMVISWKG 
               
               
                   
               
               
                 171 
                 LPWYSYLYAVSA 
               
               
                   
               
               
                 172 
                 WNLPWYYSVSPT 
               
               
                   
               
               
                 173 
                 WNL 
               
               
                   
               
               
                 174 
                 PWYYSVSPT 
               
               
                   
               
               
                 175 
                 SSWESYKSGGGTRL 
               
               
                   
               
               
                 176 
                 RDWSSQHPGRCNGETHLK 
               
               
                   
               
               
                 177 
                 SLPTLTL 
               
               
                   
               
               
                 178 
                 VICTGGDYSFALPVGQWPVMT 
               
               
                   
               
               
                 179 
                 DKPSYQFGGHNSVDFEEDTLPKV 
               
               
                   
               
               
                 180 
                 RARRRKRASATQLYQTCKASGTCPPD 
               
               
                   
               
               
                 181 
                 SGDYSFALPVGQWPWMTG 
               
               
                   
               
               
                 182 
                 CTGGDYSFALPVGQWPW 
               
               
                   
               
               
                 183 
                 FYYDYDFFFDYWGQG 
               
               
                   
               
               
                 184 
                 HLRRLRRRLLREAEG 
               
               
                   
               
               
                 185 
                 DYYCAAWDDSLNGYSVF 
               
               
                   
               
               
                 186 
                 YYCLQSMEDPYTFGG 
               
               
                   
               
               
                 187 
                 YYCARSDGNYGYYYALDYDY 
               
               
                   
               
               
                 188 
                 AARSPSYYRYDY 
               
               
                   
               
               
                 189 
                 GPYYAMDYD 
               
               
                   
               
               
                 190 
                 YYCQQRSSYPYTEGGAYPKAWK 
               
               
                   
               
               
                 191 
                 YYCQRYDSDWSFGQGTKL 
               
               
                   
               
               
                 192 
                 YYCARSGYYAMDYWGQGT 
               
               
                   
               
               
                 193 
                 RVRRGACRGDCLG 
               
               
                   
               
               
                 194 
                 RVRRGACRYDCLG 
               
               
                   
               
               
                 195 
                 YYCAKGTHWGFWSGYFDYWGQGT 
               
               
                   
               
               
                 196 
                 GRENYHGCTTHWGFTLC 
               
               
                   
               
               
                 197 
                 VQATQSNQHTPRGGGSK 
               
               
                   
               
               
                 198 
                 DPRAPGS 
               
               
                   
               
               
                 199 
                 YYCQQRSSYPYTFGG 
               
               
                   
               
               
                 200 
                 AARSPSYYRYDYGPYYAMDYD 
               
               
                   
               
               
                 201 
                 GPKLTGILISILSLFVES 
               
               
                   
               
               
                 202 
                 KYILRWRPKNS 
               
               
                   
               
               
                 203 
                 IKVAV 
               
               
                   
               
               
                 204 
                 WTPPRAQITGYRLTVGLTRR 
               
               
                   
               
               
                 205 
                 AASIKVAVSADR 
               
               
                   
               
               
                 206 
                 KLDAPT 
               
               
                   
               
               
                 207 
                 NRWHSIYITRFG 
               
               
                   
               
               
                 208 
                 PHSRN 
               
               
                   
               
               
                 209 
                 SSFHFDGSGYAM 
               
               
                   
               
               
                 210 
                 RGDS 
               
               
                   
               
               
                 211 
                 IAFQRN 
               
               
                   
               
               
                 212 
                 GRGDSP 
               
               
                   
               
               
                 213 
                 TWYKIAFQRRK 
               
               
                   
               
               
                 214 
                 EDGIHEL 
               
               
                   
               
               
                 215 
                 SLVRNRRVITIQ 
               
               
                   
               
               
                 216 
                 YRVRVTPKEKTGPMKE 
               
               
                   
               
               
                 217 
                 LQVQLSR 
               
               
                   
               
               
                 218 
                 SPPRRARVT 
               
               
                   
               
               
                 219 
                 RKRLQVQLSIRT 
               
               
                   
               
               
                 220 
                 ATETTITIS 
               
               
                   
               
               
                 221 
                 NAPFPKLSWTIQ 
               
               
                   
               
               
                 222 
                 VSPPRRARVTDATETTITISWRTKTETITGG 
               
               
                   
               
               
                 223 
                 WTIQTTVDRGLL 
               
               
                   
               
               
                 224 
                 KPDVRSYTITG 
               
               
                   
               
               
                 225 
                 DTINNGRDHMILI 
               
               
                   
               
               
                 226 
                 ANGQTPIQRYIK 
               
               
                   
               
               
                 227 
                 MILISIGKSQKRM 
               
               
                   
               
               
                 228 
                 PRARITGYIIKYEKPGSPPREVVPRPRPGV 
               
               
                   
               
               
                 229 
                 PPFLMLLKGSTR 
               
               
                   
               
               
                 230 
                 WQPPRARI 
               
               
                   
               
               
                 231 
                 NQRLASFSNAQQS 
               
               
                   
               
               
                 232 
                 WQPPRARITGYIIKYEKPG 
               
               
                   
               
               
                 233 
                 ISNVFVQRMSQSPEVLD 
               
               
                   
               
               
                 234 
                 YEKPGSPPREVVPRPRPGV 
               
               
                   
               
               
                 235 
                 KARSFNVNQLLQD 
               
               
                   
               
               
                 236 
                 KNNQKSEPLIGRKKT 
               
               
                   
               
               
                 237 
                 KNSFMALYLSKG 
               
               
                   
               
               
                 238 
                 EILDVPST 
               
               
                   
               
               
                 239 
                 KNSFMALYLSKGRLVFALG 
               
               
                   
               
               
                 240 
                 IDAPS 
               
               
                   
               
               
                 241 
                 RDSFVALYLSEGHVIFAGLG 
               
               
                   
               
               
                 242 
                 VVIDASTAIDAPSNL 
               
               
                   
               
               
                 243 
                 KPRLQFSLDIQT 
               
               
                   
               
               
                 244 
                 LDVPS 
               
               
                   
               
               
                 245 
                 DGQWHSVTVSIK 
               
               
                   
               
               
                 246 
                 REDV 
               
               
                   
               
               
                 247 
                 FVLYLGSKNAKK 
               
               
                   
               
               
                 248 
                 PHSRNRGDSP 
               
               
                   
               
               
                 249 
                 LAIKNDNLVYVY 
               
               
                   
               
               
                 250 
                 LWVTVRSQQRGLF 
               
               
                   
               
               
                 251 
                 AYFSIVKIERVG 
               
               
                   
               
               
                 252 
                 GINNWWQSPSIQN 
               
               
                   
               
               
                 253 
                 DVISLYNFKHIY 
               
               
                   
               
               
                 254 
                 WVTVTLDLRQVFQ 
               
               
                   
               
               
                 255 
                 FFDGSSYAVVRD 
               
               
                   
               
               
                 256 
                 RQVFQVAYIIIKA 
               
               
                   
               
               
                 257 
                 LHVFYDFGFGFSNG 
               
               
                   
               
               
                 258 
                 LTRYKITPRRGPPT 
               
               
                   
               
               
                 259 
                 LKKAQINDAKYREISIIYHN 
               
               
                   
               
               
                 260 
                 LLEFTSARYIRL 
               
               
                   
               
               
                 261 
                 RAYFNGQSFIAS 
               
               
                   
               
               
                 262 
                 YIRLRLQRIRTL 
               
               
                   
               
               
                 263 
                 SRLRGKNPTKGK 
               
               
                   
               
               
                 264 
                 RRYYYSIKDISV 
               
               
                   
               
               
                 265 
                 LHKKGKNSSKPK 
               
               
                   
               
               
                 266 
                 SINNTAVNQRLT 
               
               
                   
               
               
                 267 
                 RLKTRSSHGMIF 
               
               
                   
               
               
                 268 
                 GGFLKYTVSYDI 
               
               
                   
               
               
                 269 
                 GEKSQFSIRLKT 
               
               
                   
               
               
                 270 
                 RDQLMTVLANVT 
               
               
                   
               
               
                 271 
                 TLFLAHGRLVFM 
               
               
                   
               
               
                 272 
                 ANVTHLLIRANY 
               
               
                   
               
               
                 273 
                 LVFMFNVGHKKL 
               
               
                   
               
               
                 274 
                 AGTFALRGDNPQG 
               
               
                   
               
               
                 275 
                 TLFLAHGRLVFMFNVGHKKL 
               
               
                   
               
               
                 276 
                 VLIKGGRARKHV 
               
               
                   
               
               
                 277 
                 DFMTLFLAHGRLVFMGNVG 
               
               
                   
               
               
                 278 
                 LSNIDYLIKAS 
               
               
                   
               
               
                 279 
                 HKKLKIRSQEKY 
               
               
                   
               
               
                 280 
                 LQQSRIANISME 
               
               
                   
               
               
                 281 
                 GAAWKIKGPIYL 
               
               
                   
               
               
                 282 
                 NLLLLLVKANLK 
               
               
                   
               
               
                 283 
                 VIRDSNVVQLDV 
               
               
                   
               
               
                 284 
                 HRDELLLWARKI 
               
               
                   
               
               
                 285 
                 GLIYYVAHQNQM 
               
               
                   
               
               
                 286 
                 KRRARDLVHRAE 
               
               
                   
               
               
                 287 
                 DYATLQLQEGRLHFMFDLG 
               
               
                   
               
               
                 288 
                 SQFQESVDNITK 
               
               
                   
               
               
                 289 
                 KKGSYNNIVVHV 
               
               
                   
               
               
                 290 
                 PGGMREKGRKAR 
               
               
                   
               
               
                 291 
                 ADNLLFYLGSAK 
               
               
                   
               
               
                 292 
                 MEMQANLLLDRL 
               
               
                   
               
               
                 293 
                 GSAKFIDFLAIE 
               
               
                   
               
               
                 294 
                 LSEIKLLISAR 
               
               
                   
               
               
                 295 
                 KVSFLWWVGSGV 
               
               
                   
               
               
                 296 
                 RDFTKATNIRLRFLR 
               
               
                   
               
               
                 297 
                 SYWYRIEASRTG 
               
               
                   
               
               
                 298 
                 ISTVMFKFRTFS 
               
               
                   
               
               
                 299 
                 YFDGTGFAKAVG 
               
               
                   
               
               
                 300 
                 KQANISIVDIDSN 
               
               
                   
               
               
                 301 
                 NGQWHKVTAKKI 
               
               
                   
               
               
                 302 
                 FSTRNESGIILL 
               
               
                   
               
               
                 303 
                 AKKIKNRLELVV 
               
               
                   
               
               
                 304 
                 RRQTTQAYYAIF 
               
               
                   
               
               
                 305 
                 GFPGGLNQFGLTTN 
               
               
                   
               
               
                 306 
                 YAIFLNKGRLEV 
               
               
                   
               
               
                 307 
                 NQFGLTTNIRFRG 
               
               
                   
               
               
                 308 
                 KNRLTIELEVRT 
               
               
                   
               
               
                 309 
                 IRSLKLTKGTGKP 
               
               
                   
               
               
                 310 
                 GLLFYMARINHA 
               
               
                   
               
               
                 311 
                 AKALELRGVQPVS 
               
               
                   
               
               
                 312 
                 VQLRNGFPYFSY 
               
               
                   
               
               
                 313 
                 GQLFHVAYILIKF 
               
               
                   
               
               
                 314 
                 HKIKIVRVKQEG 
               
               
                   
               
               
                 315 
                 NVLSLYNFKTTF 
               
               
                   
               
               
                 316 
                 DFGTVQLRNGFPFFSYDLG 
               
               
                   
               
               
                 317 
                 SQRIYQFAKLNYT 
               
               
                   
               
               
                 318 
                 NIRLRFLRTNTL 
               
               
                   
               
               
                 319 
                 EVNVTLDLGQVFH 
               
               
                   
               
               
                 320 
                 GKNTGDHFVLYM 
               
               
                   
               
               
                 321 
                 GQVFHVAYVLIKF 
               
               
                   
               
               
                 322 
                 VVSLYNFEQTFML 
               
               
                   
               
               
                 323 
                 HQQDLGTAGSCLRKFSTMFLF 
               
               
                   
               
               
                 324 
                 RFDQELRLVSYN 
               
               
                   
               
               
                 325 
                 HQQDLGTAGSCLRKFSTMFLFCNI 
               
               
                   
               
               
                 326 
                 RLVSYSGVLFFLK 
               
               
                   
               
               
                 327 
                 VAEIDGIEL 
               
               
                   
               
               
                 328 
                 NWRHISYITRFG 
               
               
                   
               
               
                 329 
                 GIIFFL 
               
               
                   
               
               
                 330 
                 KRLQVQLRSIRT 
               
               
                   
               
               
                 331 
                 ASKAIQVFLLGG 
               
               
                   
               
               
                 332 
                 TWYKIAFQRNRK 
               
               
                   
               
               
                 333 
                 VLVRVERATVFS 
               
               
                   
               
               
                 334 
                 QVFQVAYIIIKA 
               
               
                   
               
               
                 335 
                 TVFSVDQDNMLE 
               
               
                   
               
               
                 336 
                 GEFYFDLRLKGDK 
               
               
                   
               
               
                 337 
                 RLRGPQRVFDLH 
               
               
                   
               
               
                 338 
                 GTPGPQGIA 
               
               
                   
               
               
                 339 
                 FDLHQNMGSVN 
               
               
                   
               
               
                 340 
                 GQRDVV 
               
               
                   
               
               
                 341 
                 LRAHAVDVNG 
               
               
                   
               
               
                 342 
                 TAGSCLRKFSTM 
               
               
                   
               
               
                 343 
                 LFSHAVSSNG 
               
               
                   
               
               
                 344 
                 KGHRGF 
               
               
                   
               
               
                 345 
                 TAGSCLRKFSTMFLF 
               
               
                   
               
               
                 346 
                 TAGSCLRKFSTMFLFCNI 
               
               
                   
               
               
                 347 
                 DLGTAGSCLRKFSTM 
               
               
                   
               
               
                 348 
                 HQQDLGTAGSCLRKFSTM 
               
               
                   
               
               
                 349 
                 RNIAEIIKDI 
               
               
                   
               
               
                 350 
                 SIGFRGDGQTC 
               
               
                   
               
               
                 351 
                 LNRQELFPFG 
               
               
                   
               
               
                 352 
                 RIQNLLKITNLRIKFVK 
               
               
                   
               
               
                 353 
                 KKQRFRHRNRKGYRSQ 
               
               
                   
               
               
                 354 
                 SINNTAVMQRLT 
               
               
                   
               
               
                 355 
                 FRHRNRKGY 
               
               
                   
               
               
                 356 
                 RYRVRVTPKEKTGPMKE 
               
               
                   
               
               
                 357 
                 SETTVKYIFRLHE 
               
               
                   
               
               
                 358 
                 GHRGPTGRPGKRGKQGQKGDS 
               
               
                   
               
               
                 359 
                 KAFDITYVRLKF 
               
               
                   
               
               
                 360 
                 GDLGRPGRKGRPGPP 
               
               
                   
               
               
                 361 
                 YIGSR 
               
               
                   
               
               
                 362 
                 RGEFYFDLRLKGDK 
               
               
                   
               
               
                 363 
                 LAGSCLARFSTM 
               
               
                   
               
               
                 364 
                 LALFLSNGHFVA 
               
               
                   
               
               
                 365 
                 ISRCQVCMKKRH 
               
               
                   
               
               
                 366 
                 PGRWHKVSVRWE 
               
               
                   
               
               
                 367 
                 TDIPPCPHGWISLWK 
               
               
                   
               
               
                 368 
                 VRWGMQQIQLVV 
               
               
                   
               
               
                 369 
                 TAIPSCPEGTVPLYS 
               
               
                   
               
               
                 370 
                 KMPYVSLELEMR 
               
               
                   
               
               
                 371 
                 GPAGKDGEAGAQG 
               
               
                   
               
               
                 372 
                 VLLQANDGAGEF 
               
               
                   
               
               
                 373 
                 GLPGER 
               
               
                   
               
               
                 374 
                 DGRWHRVAVIMG 
               
               
                   
               
               
                 375 
                 LAGSCLPVFSTL 
               
               
                   
               
               
                 376 
                 APVNVTASVQIQ 
               
               
                   
               
               
                 377 
                 TAGSCLRRFSTM 
               
               
                   
               
               
                 378 
                 KQGKALTQRHAK 
               
               
                   
               
               
                 379 
                 TAGSCLRKF 
               
               
                   
               
               
                 380 
                 RYVVLPR 
               
               
                   
               
               
                 381 
                 TAGSCL 
               
               
                   
               
               
                 382 
                 SPYTFIDSLVLMPY 
               
               
                   
               
               
                 383 
                 TAG 
               
               
                   
               
               
                 384 
                 PDSGR 
               
               
                   
               
               
                 385 
                 QQNLGSVNVSTG 
               
               
                   
               
               
                 386 
                 SRATAQKVSRRS 
               
               
                   
               
               
                 387 
                 DPGYIGSR 
               
               
                   
               
               
                 388 
                 GSLSSHLEFVGI 
               
               
                   
               
               
                 389 
                 VILQQSAADIAR 
               
               
                   
               
               
                 390 
                 RNRLHLSMLVRP 
               
               
                   
               
               
                 391 
                 KDISEKVAVYST 
               
               
                   
               
               
                 392 
                 APMSGRSPSLVLK 
               
               
                   
               
               
                 393 
                 LGTIPG 
               
               
                   
               
               
                 394 
                 AFGVLALWGTRV 
               
               
                   
               
               
                 395 
                 TDIRVTLNRLNTF 
               
               
                   
               
               
                 396 
                 IENVVTTFAPNR 
               
               
                   
               
               
                 397 
                 AFSTLEGRPSAY 
               
               
                   
               
               
                 398 
                 LEAEFHFTHLIM 
               
               
                   
               
               
                 399 
                 TSAEAYNLLLRT 
               
               
                   
               
               
                 400 
                 HLIMTFKTFRPA 
               
               
                   
               
               
                 401 
                 LNRRYEQARNIS 
               
               
                   
               
               
                 402 
                 KTWGVYRYFAYD 
               
               
                   
               
               
                 403 
                 SLLSQLNNLLDQ 
               
               
                   
               
               
                 404 
                 TNLRIKFVKLHT 
               
               
                   
               
               
                 405 
                 RDIAEIIKDI 
               
               
                   
               
               
                 406 
                 KRLVTGQR 
               
               
                   
               
               
                 407 
                 SHAVSS 
               
               
                   
               
               
                 408 
                 GPGVVVVERQYI 
               
               
                   
               
               
                 409 
                 ADTPPV 
               
               
                   
               
               
                 410 
                 NEPKVLKSYYYAI 
               
               
                   
               
               
                 411 
                 LRAHAVDING 
               
               
                   
               
               
                 412 
                 YYAISDFAVGGR 
               
               
                   
               
               
                 413 
                 DSITKYFQMSLE 
               
               
                   
               
               
                 414 
                 LPFFNDRPWRRAT 
               
               
                   
               
               
                 415 
                 YTALIIATDN 
               
               
                   
               
               
                 416 
                 FDPELYRSTGHGGH 
               
               
                   
               
               
                 417 
                 VITVKDINDN 
               
               
                   
               
               
                 418 
                 TNAVGYSVYDIS 
               
               
                   
               
               
                 419 
                 GLDRESYPYY 
               
               
                   
               
               
                 420 
                 APVKFLGNQVLSY 
               
               
                   
               
               
                 421 
                 MKVSATDADD 
               
               
                   
               
               
                 422 
                 SFSFRVDRRDTR 
               
               
                   
               
               
                 423 
                 PQVTRGDVFTMP 
               
               
                   
               
               
                 424 
                 TWSKVGGHLRPGIVQSG 
               
               
                   
               
               
                 425 
                 KEAEREVTDLLR 
               
               
                   
               
               
                 426 
                 RGDV 
               
               
                   
               
               
                 427 
                 AAEPLKNIGILF 
               
               
                   
               
               
                 428 
                 FALWDAIIGEL 
               
               
                   
               
               
                 429 
                 VGVAPG 
               
               
                   
               
               
                 430 
                 LWPLLAVLAAVA 
               
               
                   
               
               
                 431 
                 PGVGV 
               
               
                   
               
               
                 432 
                 VFDNFVLK 
               
               
                   
               
               
                 433 
                 TSIKIRGTYSER 
               
               
                   
               
               
                 434 
                 TTSWSQCSKS 
               
               
                   
               
               
                 435 
                 DPETGV 
               
               
                   
               
               
                 436 
                 KRSR 
               
               
                   
               
               
                 437 
                 QGADTPPVGV 
               
               
                   
               
               
                 438 
                 SVVYGLR 
               
               
                   
               
               
                 439 
                 PLDREAIAKY 
               
               
                   
               
               
                 440 
                 DGRGDSVAYG 
               
               
                   
               
               
                 441 
                 HAVDI 
               
               
                   
               
               
                 442 
                 LALERKDHSG 
               
               
                   
               
               
                 443 
                 DQNDN 
               
               
                   
               
               
                 444 
                 YSMKKTTMKIIPFNRLTIG 
               
               
                   
               
               
                 445 
                 QDPELPDKNM 
               
               
                   
               
               
                 446 
                 RGDF 
               
               
                   
               
               
                 447 
                 LVVQAADLQG 
               
               
                   
               
               
                 448 
                 GVYYQGGTYSKAS 
               
               
                   
               
               
                 449 
                 NDDGGQFVVT 
               
               
                   
               
               
                 450 
                 TAGSCLRKFSCL 
               
               
                   
               
               
                 451 
                 YILHVAVTN 
               
               
                   
               
               
                 452 
                 CNYYSNSYSFWLASLNPER 
               
               
                   
               
               
                 453 
                 TYRIWRDTAN 
               
               
                   
               
               
                 454 
                 TGLSCLQRFTTM 
               
               
                   
               
               
                 455 
                 GFTCECSIGFRGDGQTCYGIVFWSEV 
               
               
                   
               
               
                 456 
                 HHLGGAKQAGDV 
               
               
                   
               
               
                 457 
                 SCLPGFSGDGRACRDVDECGH 
               
               
                   
               
               
                 458 
                 MAPRPSLAKKQRFRHRNRKGYRSQRGHSRG 
               
               
                   
               
               
                 459 
                 KKQKFRHRNRKGYRSQ 
               
               
                   
               
               
                 460 
                 KKQKFKHRNRKGYRS 
               
               
                   
               
               
                 461 
                 KKQKFRRRNRKGYRSH 
               
               
                   
               
               
                 462 
                 TAIPPCPHGWISLWK 
               
               
                   
               
               
                 463 
                 KKQKSRHRSRKRYRS 
               
               
                   
               
               
                 464 
                 KKQKSRRRSRKGYRS 
               
               
                   
               
               
                 465 
                 ISRCTVC 
               
               
                   
               
               
                 466 
                 ISRCQVCMKRRH 
               
               
                   
               
               
                 467 
                 VSRCTVC 
               
               
                   
               
               
                 468 
                 TDIPPCPQGWISLWK 
               
               
                   
               
               
                 469 
                 TVKAGELEKIISRCQVMKKRH 
               
               
                   
               
               
                 470 
                 TDIPSCPHGWISLWK 
               
               
                   
               
               
                 471 
                 TDIPPCPAGWISLWK 
               
               
                   
               
               
                 472 
                 TEIPPCPQGWISLWK 
               
               
                   
               
               
                 473 
                 TDVPPCPQGWISLWK 
               
               
                   
               
               
                 474 
                 RLVSYNGILFFLK 
               
               
                   
               
               
                 475 
                 RLVSYSGVIFFLK 
               
               
                   
               
               
                 476 
                 RLVSYNGILFFL 
               
               
                   
               
               
                 477 
                 RLVSYSGIIFFLK 
               
               
                   
               
               
                 478 
                 RFEQELRLVSYSGVLFFLKQ 
               
               
                   
               
               
                 479 
                 RLVSYNGIIFFLK 
               
               
                   
               
               
                 480 
                 DPAFKIEDPYSPRIQNLLKITNLRIKFVKL 
               
               
                   
               
               
                 481 
                 TKRFEQELRLVSYSGVLFFL 
               
               
                   
               
               
                 482 
                 GGRLKYSVAF 
               
               
                   
               
               
                 483 
                 GGFLRYTVSYDI 
               
               
                   
               
               
                 484 
                 GGFLKYTVSYDV 
               
               
                   
               
               
                 485 
                 LGNKLTAFGGFLKYTVSYDIPV 
               
               
                   
               
               
                 486 
                 GGYLKYTVSYDI 
               
               
                   
               
               
                 487 
                 GEIFFDMRLKGDK 
               
               
                   
               
               
                 488 
                 GEIYFDLRLKGDK 
               
               
                   
               
               
                 489 
                 GEIYLDMRLKGDK 
               
               
                   
               
               
                 490 
                 IGQPGAKGEPGEFYFDLRLKGDKGDPGFPG 
               
               
                   
               
               
                 491 
                 GEVFFDMRLKGDK 
               
               
                   
               
               
                 492 
                 LAGSCLPIFSTL 
               
               
                   
               
               
                 493 
                 AHNQDLGLAGSCLARFSTMPFLYCNPGDIC 
               
               
                   
               
               
                 494 
                 QEKAHNQDLGLAGSCLPVFSTLPFAYCNIH 
               
               
                   
               
               
                 495 
                 LAGSCLPVFSTM 
               
               
                   
               
               
                 496 
                 GNKRAHGQDLGTAGSCLRRFSTMPFNIFCNI 
               
               
                   
               
               
                 497 
                 RAHGQDLGTAGSCLRRFSTMP 
               
               
                   
               
               
                 498 
                 RKRLQVQLNIRT 
               
               
                   
               
               
                 499 
                 HLVLPLQQSDVRKRLQVQLSIRTFASSGLI 
               
               
                   
               
               
                 500 
                 RKRLSVQLRIRT 
               
               
                   
               
               
                 501 
                 DLGTAGSCLRRFSTM 
               
               
                   
               
               
                 502 
                 RNIAEIIKDI 
               
               
                   
               
               
                 503 
                 TAGSCLRKFSTMRRRRRRRRRRRR 
               
               
                   
               
               
                 504 
                 FTLTGLLGTLVTMGLLT 
               
               
                   
               
               
                 505 
                 APYKAWK 
               
               
                   
               
               
                 506 
                 STSKTNRGDDSNWSKRVTNNKPS 
               
               
                   
               
               
                 507 
                 STSKRKRGDDSNWSKRVTKKKPS 
               
               
                   
               
               
                 508 
                 STSKRKRGDDSNWSKRVSKKKPS 
               
               
                   
               
               
                 509 
                 STSKRKRGDDANWSKRVTKKKPS 
               
               
                   
               
               
                 510 
                 PLAGSKRKRADEVAWSKRGTKKKPER 
               
               
                   
               
               
                 511 
                 PLAGSKRKRADEVAWSKRGTKKKPERTSAARAGPSRRIR 
               
               
                   
               
               
                 512 
                 STSKRKRGDDANWSKRTTKKKPSS 
               
               
                   
               
               
                 513 
                 STSKRKRGDDANWSKRTTKKKPSSAGLKRAGSKADRPSL 
               
               
                   
               
               
                 514 
                 PTTAGKRKRSDDAAWSKRARPKAGRT 
               
               
                   
               
               
                 515 
                 PTTAGKRKRSDDAAWSKRARPKAGRTSAARPGTSVRRIR 
               
               
                   
               
               
                 516 
                 SSSLGKRKRSDEGAWSKGKSKKKAMR 
               
               
                   
               
               
                 517 
                 SSSLGKRKRSDEGAWSKGKSKKKAMRGSSSRRPGPVRGP 
               
               
                   
               
               
                 518 
                 PTTAGKRKRTDDAAWSKRARPKAGR 
               
               
                   
               
               
                 519 
                 PTTAGKRKRTDDAAWSKRARPKAGRTSAARPGTAVRRVR 
               
               
                   
               
               
                 520 
                 PATAGKRKRSDDAAWSKRARPKAGRTSAAR 
               
               
                   
               
               
                 521 
                 PATAGKRKRSDDAAWSKRARPKAGRTSAARPGTSVRRIR 
               
               
                   
               
               
                 522 
                 SSSLGKRKRSNGGDWSKRSAKKKPA 
               
               
                   
               
               
                 523 
                 SSSLGKRKRSNGGDWSKRSAKKKPAGTPSRRAGPGRGPR 
               
               
                   
               
               
                 524 
                 SSSLGKRKRSDEGAWSKGKSKKKAMR 
               
               
                   
               
               
                 525 
                 SSSLGKRKRSDEGAWSKGKSKKKAMRGSSSRRPGPVRGP 
               
               
                   
               
               
                 526 
                 STSKRKRGDDANWNKRPTKKKPSS 
               
               
                   
               
               
                 527 
                 STSKRKRGDDANWNKRPTKKKPSSAGLKKAGSKAERPSL 
               
               
                   
               
               
                 528 
                 SGALKRKRSDEVAWSRRRPVKKPV 
               
               
                   
               
               
                 529 
                 SGALKRKRSDEVAWSRRRPVKKPVRRAPPPRAGPSVRRG 
               
               
                   
               
               
                 530 
                 SGALKRKRSDEVAWSRRKPAKKPAR 
               
               
                   
               
               
                 531 
                 SGALKRKRSDEVAWSRRKPAKKPARQPPPPRAGPSVRRG 
               
               
                   
               
               
                 532 
                 AGALKRKRSDEVAWSRRKPAKKPAR 
               
               
                   
               
               
                 533 
                 AGALKRKRSDEVAWSRRKPAKKPARAPPPRAGPSVRRGL 
               
               
                   
               
               
                 534 
                 STSKRKRGDDSNWSKRVTKKKPSSAGLKRAGSKADRPSL 
               
               
                   
                 QIQTLQHAGTTMITVPSGGVCDLINTYARGSDEGNRHTS 
               
               
                   
                 ETLTYKIAIDYHFVADAAACRYSNTGTGVMWLVYDTTPG 
               
               
                   
                 GQAPTPQTIFSYPDTLKAWPATWKVSRELCHRFVVKRRW 
               
               
                   
                 LFNMETDGRIGSDIPPSNASWKPCKRNIYFHKFTSGLGV 
               
               
                   
                 RTQWKNVTDGGVGAIQRGALYMVIAPGNGLTFTAHGQTR 
               
               
                   
                 LYFKSVGNQ 
               
               
                   
               
               
                 535 
                 DPQNALYYQPRVPTAAPTSGGVPWSRVGEVAILSFVALI 
               
               
                   
                 CFYLLYLWVLRDLILVLKARQGRSTEELIFGGQAVDRSN 
               
               
                   
                 PIPNIPAPPSQGNPGPFVPGTG 
               
               
                   
               
               
                 536 
                 GSQLVPPPSAFNYIESQRDEFQLSHDLTEIVLQFPSTAS 
               
               
                   
                 QITARLSRSCMKIDHCVIEYRQQVPINASGTVIVEIHDK 
               
               
                   
                 RMTDNESLQASWTFPIRCNIDLHYFSSSFFSLKDPIPWK 
               
               
                   
                 LYYRVSDSNVHQMTHFAKFKGKLKLSSAKHSVDIPFRAP 
               
               
                   
                 TVKILAKQFSEKDIDFWHVGYGKWERRLVKSASSSRFGL 
               
               
                   
                 RGPIEINPGESWATKSAIVTPNRNADLDIEEELLPYREL 
               
               
                   
                 NRLGTNILDPGESASIVGIQRSQSNITMSMSQLNELVRS 
               
               
                   
                 TVHECIKTSCIPSTPKSLS 
               
               
                   
               
               
                 537 
                 RTGVKRSYGAARGDDRRRPNVV 
               
               
                   
               
               
                 538 
                 SYVKTVPNRTRTYIKLRVR 
               
               
                   
               
               
                 539 
                 MYSTSNRRGRSQTQRGSHVRRTGVKRSYGAARGDDRRRP 
               
               
                   
                 NVVSKTQVEPRMTIQRVQENQFGPEFVLSQNSALSTFVT 
               
               
                   
                 YPSYVKTVPNRTRTYIKLKRVRFKGTLKIERGQGDTIMD 
               
               
                   
                 GPSSNIEGVFSMVIVVDRKPHVSQSGRLHTFDELFGARI 
               
               
                   
                 HCHGNLSVVPALKDRYYIRHVTKRVVSLEKDTLLIDLHG 
               
               
                   
                 TTQLSNKRYNCWASFSDLERDCNGVYGNITKNALLVYYC 
               
               
                   
                 WLSDAQSKASTYVSFELDYLG 
               
               
                   
               
               
                 540 
                 R 12 -VDYGKWERKPIRCASMSR 
               
               
                   
               
               
                 541 
                 R 12 -GKWERKPIRCAS 
               
               
                   
               
               
                 542 
                 K 16 -GKWERKPIRCAS 
               
               
                   
               
               
                 543 
                 R 12 -VDFSHVDYGKWERKPIRCASMSRLGLRG 
               
               
                   
               
               
                 544 
                 GVKRSYGAARGDDRRRPNVVAPYKAW-R 12   
               
               
                   
               
               
                 545 
                 KSVPNRTRTYIKLKRLRFKGAPYKAW-R 12   
               
               
                   
               
               
                 546 
                 RTGVKRSYGAARGDDRRRPNVV-R 12   
               
               
                   
               
               
                 547 
                 SYVKTVPNRTRTYIKGGGGG-R 12   
               
               
                   
               
               
                 548 
                 VDIPFRAPTIKILSKQFTEDDIDFWHVGYGKWERKLVRPA 
               
               
                   
                 SLSGRRGLRR 
               
               
                   
               
               
                 549 
                 IDFWHVGYGKWERKLVRPASLSGRRGLRR 
               
               
                   
               
               
                 550 
                 IDFWSVEKGETRRRLLNPTPHAHSPRPIAHR 
               
               
                   
               
               
                 551 
                 IDFSHVGYGKWERKMIRSASISRLGLHN 
               
               
                   
               
               
                 552 
                 VDFSHVGYGKWERKLIRSASTVKYGLPS 
               
               
                   
               
               
                 553 
                 IDFSHVDYGKVERKLVKCESSSRLGLHS 
               
               
                   
               
               
                 554 
                 IDFWSVGRKAQQRKLVQGPSLIGSRSMRY 
               
               
                   
               
               
                 555 
                 IDFWSVGSKPQTRRLVDGSRLIGHSSRSLRV 
               
               
                   
               
               
                 556 
                 IDFWSVERGETRRRLLNPTPSAGSNRALSKR 
               
               
                   
               
               
                 557 
                 VDFWSVGKPKPIRRLIQNDPGTDYDTGPKYR 
               
               
                   
               
               
                 558 
                 VDFWSVEKPKPIRRLLNPGPNQGPYPNTGHR 
               
               
                   
               
               
                 559 
                 VDFSHVDYGKWERKLIRSASTSRYGLRS 
               
               
                   
               
               
                 560 
                 VDFSHVDYGKWERKTLRSRSLSRIGLTG 
               
               
                   
               
               
                 561 
                 IDFWHVGYGKWERRLVKSASSSRFGIRG 
               
               
                   
               
               
                 562 
                 VDFFHVDYGRWERKHIRCASMSRVGLRG 
               
               
                   
               
               
                 563 
                 GTFQHVDYGKWERKPIRCQSMSRVGYRR 
               
               
                   
               
               
                 564 
                 VGYGKWERKLVRPASLS 
               
               
                   
               
               
                 565 
                 VEKGETRRRLLNPTPHA 
               
               
                   
               
               
                 566 
                 VGYGKWERKLIRSASTV 
               
               
                   
               
               
                 567 
                 VEKPKPIRRLLNPGPNQ 
               
               
                   
               
               
                 568 
                 VDYGKWERKLIRSASTS 
               
               
                   
               
               
                 569 
                 VDYGKWERKTLRSRSLS 
               
               
                   
               
               
                 570 
                 VGYGKWERRLVKSASSS 
               
               
                   
               
               
                 571 
                 VDYGRWERKHIRCASMS 
               
               
                   
               
               
                 572 
                 VERPKPIRRLLTPTPGC 
               
               
                   
               
               
                 573 
                 PFRAPTIKILSKQFTEDDIDFWHVGYGKWERKLVRPAS 
               
               
                   
                 LSGRRGLRR 
               
               
                   
               
               
                 574 
                 PFRAPTVKILSKQFTDKDIDFSHVGYGKWERKMIRSA 
               
               
                   
                 SISRLGL 
               
               
                   
               
               
                 575 
                 DIAFRAPTVKILSKQFTDRDVDFSHVGYGKWERKLIR 
               
               
                   
                 SASTVKYGL 
               
               
                   
               
               
                 576 
                 DIRFKPPTINILSKDYTADCVDFWSVEKPKPIRRLLNPG 
               
               
                   
                 PNQGPYPNTG 
               
               
                   
               
               
                 577 
                 DIPFRAPTVKIHSKQFSHRDVDFSHVDYGKWERKTLRSR 
               
               
                   
                 SLSRIGL 
               
               
                   
               
               
                 578 
                 DIPFRAPTVKILAKQFSEKDIDFWHVGYGKWERRLVKSA 
               
               
                   
                 SSSRFGI 
               
               
                   
               
               
                 579 
                 DIPFRAPTVKILSKQFTDKDVDFFHVDYGRWERKHIRCA 
               
               
                   
                 SMSRVGL 
               
               
                   
               
               
                 580 
                 DIKYKPPTIKILSKDYTADCVDFWSVERPKPIRRLLTPTPGCG 
               
               
                   
               
               
                 581 
                 ARTKQTARKSTGGKAPRKQLATKAARKSAPATGGVK 
               
               
                   
                 KPHRYRPGTVA 
               
               
                   
               
               
                 582 
                 SGRGKGGKGLGKGGAKRHRKVLRDNIQGITKPAI 
               
               
                   
               
            
           
         
       
     
     
       
         
           
               
             
               
                 TABLE 2 
               
             
            
               
                   
               
               
                 Relative Fluorescence of Transfection of HeLa Cells 
               
            
           
           
               
               
               
               
               
            
               
                   
                 SEQ ID NO 
                 RF* 
                 SEQ ID NO 
                 RF 
               
               
                   
                   
               
            
           
           
               
               
               
               
               
            
               
                   
                 DOTMA 
                 7551 
                 327 
                 5345 
               
               
                   
                 107 
                 75 
                 328 
                 32528 
               
               
                   
                 205 
                 26368 
                 332 
                 9198 
               
               
                   
                 216 
                 6181 
                 335 
                 26954 
               
               
                   
                 218 
                 11179 
                 336 
                 25262 
               
               
                   
                 219 
                 13721 
                 338 
                 6241 
               
               
                   
                 220 
                 9398 
                 341 
                 4108 
               
               
                   
                 224 
                 5559 
                 342 
                 33666 
               
               
                   
                 226 
                 7340 
                 343 
                 5910 
               
               
                   
                 229 
                 8816 
                 344 
                 14415 
               
               
                   
                 230 
                 8944 
                 345 
                 8493 
               
               
                   
                 234 
                 3867 
                 347 
                 22527 
               
               
                   
                 236 
                 7202 
                 348 
                 22289 
               
               
                   
                 236 
                 5042 
                 349 
                 4433 
               
               
                   
                 237 
                 21288 
                 350 
                 13313 
               
               
                   
                 238 
                 6067 
                 351 
                 12279 
               
               
                   
                 239 
                 2717 
                 352 
                 32126 
               
               
                   
                 256 
                 2243 
                 353 
                 29119 
               
               
                   
                 268 
                 25885 
                 354 
                 4403 
               
               
                   
                 323 
                 14067 
                 355 
                 32628 
               
               
                   
                 326 
                 18946 
                 357 
                 22679 
               
               
                   
                   
                   
                 358 
                 15132 
               
               
                   
                   
                   
                 359 
                 3136 
               
               
                   
                   
                   
                 360 
                 8516 
               
               
                   
                   
                   
                 361 
                 3698 
               
               
                   
                   
                   
                 363 
                 2590 
               
               
                   
                   
                   
                 365 
                 36093 
               
               
                   
                   
                   
                 367 
                 39008 
               
               
                   
                   
                   
                 369 
                 14444 
               
               
                   
                   
                   
                 371 
                 5180 
               
               
                   
                   
                   
                 373 
                 8992 
               
               
                   
                   
                   
                 375 
                 41076 
               
               
                   
                   
                   
                 377 
                 15091 
               
               
                   
                   
                   
                 379 
                 27424 
               
               
                   
                   
                   
                 381 
                 22652 
               
               
                   
                   
                   
                 383 
                 8012 
               
               
                   
                   
                   
                 450 
                 21045 
               
               
                   
                   
                   
                 452 
                 13888 
               
               
                   
                   
                   
                 454 
                 34550 
               
               
                   
                   
                   
                 503 
                 25368 
               
               
                   
                   
               
               
                   
                 *RF = Relative Fluorescence 
               
            
           
         
       
     
     
       
         
           
               
             
               
                 TABLE 3 
               
             
            
               
                   
               
               
                 Relative Fluorescence of Transfection of HuVec Cells 
               
            
           
           
               
               
               
               
               
            
               
                   
                 SEQ ID NO 
                 RF* 
                 SEQ ID NO 
                 RF 
               
               
                   
                   
               
            
           
           
               
               
               
               
               
            
               
                   
                 DOTMA 
                 12230 
                 328 
                 3614 
               
               
                   
                 205 
                 8309 
                 332 
                 4221 
               
               
                   
                 216 
                 14693 
                 335 
                 6350 
               
               
                   
                 218 
                 14827 
                 336 
                 5366 
               
               
                   
                 219 
                 6237 
                 338 
                 12000 
               
               
                   
                 220 
                 8737 
                 341 
                 15906 
               
               
                   
                 224 
                 15945 
                 342 
                 12804 
               
               
                   
                 226 
                 20864 
                 343 
                 16239 
               
               
                   
                 229 
                 5452 
                 344 
                 11867 
               
               
                   
                 230 
                 9067 
                 345 
                 2939 
               
               
                   
                 234 
                 13956 
                 347 
                 20728 
               
               
                   
                 235 
                 26940 
                 348 
                 21214 
               
               
                   
                 236 
                 14418 
                 349 
                 22907 
               
               
                   
                 237 
                 2302 
                 350 
                 13163 
               
               
                   
                 238 
                 11083 
                 351 
                 15379 
               
               
                   
                 239 
                 1404 
                 352 
                 3130 
               
               
                   
                 256 
                 3030 
                 353 
                 8772 
               
               
                   
                 268 
                 5403 
                 354 
                 19284 
               
               
                   
                 323 
                 12815 
                 355 
                 14810 
               
               
                   
                 326 
                 4932 
                 357 
                 6009 
               
               
                   
                 327 
                 21018 
                 358 
                 25398 
               
               
                   
                   
                   
                 359 
                 3024 
               
               
                   
                   
                   
                 360 
                 24042 
               
               
                   
                   
                   
                 361 
                 6017 
               
               
                   
                   
                   
                 363 
                 1383 
               
               
                   
                   
                   
                 365 
                 14764 
               
               
                   
                   
                   
                 367 
                 13354 
               
               
                   
                   
                   
                 369 
                 6536 
               
               
                   
                   
                   
                 371 
                 10669 
               
               
                   
                   
                   
                 373 
                 28811 
               
               
                   
                   
                   
                 375 
                 5542 
               
               
                   
                   
                   
                 377 
                 9326 
               
               
                   
                   
                   
                 379 
                 14573 
               
               
                   
                   
                   
                 381 
                 17542 
               
               
                   
                   
                   
                 383 
                 20267 
               
               
                   
                   
                   
                 450 
                 20911 
               
               
                   
                   
                   
                 452 
                 1841 
               
               
                   
                   
                   
                 454 
                 13336 
               
               
                   
                   
                   
                 503 
                 12123 
               
               
                   
                   
               
               
                   
                 *RF = Relative Fluorescence 
               
            
           
         
       
     
     
       
         
           
               
             
               
                 TABLE 4 
               
             
            
               
                   
               
               
                 Relative Fluorescence of Transfection of NL-1 iPS Cells 
               
            
           
           
               
               
               
               
               
            
               
                   
                 SEQ ID NO 
                 RF* 
                 SEQ ID NO 
                 RF 
               
               
                   
                   
               
            
           
           
               
               
               
               
               
            
               
                   
                 DOTMA 
                 8965 
                 327 
                 31383 
               
               
                   
                 107 
                 1441 
                 328 
                 8611 
               
               
                   
                 205 
                 13140 
                 332 
                 8725 
               
               
                   
                 216 
                 19626 
                 335 
                 14424 
               
               
                   
                 218 
                 17049 
                 336 
                 14875 
               
               
                   
                 219 
                 5770 
                 338 
                 17288 
               
               
                   
                 220 
                 15892 
                 341 
                 20446 
               
               
                   
                 224 
                 25209 
                 342 
                 11493 
               
               
                   
                 226 
                 24920 
                 343 
                 21726 
               
               
                   
                 229 
                 13491 
                 344 
                 10895 
               
               
                   
                 230 
                 14524 
                 345 
                 7836 
               
               
                   
                 234 
                 14410 
                 347 
                 21865 
               
               
                   
                 236 
                 43542 
                 348 
                 21645 
               
               
                   
                 236 
                 22256 
                 349 
                 33103 
               
               
                   
                 237 
                 8632 
                 350 
                 19132 
               
               
                   
                 238 
                 15856 
                 351 
                 24646 
               
               
                   
                 239 
                 4129 
                 352 
                 11628 
               
               
                   
                 256 
                 1825 
                 353 
                 9248 
               
               
                   
                 268 
                 15697 
                 354 
                 24043 
               
               
                   
                 323 
                 21694 
                 355 
                 7910 
               
               
                   
                 326 
                 9239 
                 357 
                 15624 
               
               
                   
                   
                   
                 358 
                 21956 
               
               
                   
                   
                   
                 359 
                 5548 
               
               
                   
                   
                   
                 360 
                 27572 
               
               
                   
                   
                   
                 361 
                 9363 
               
               
                   
                   
                   
                 363 
                 4189 
               
               
                   
                   
                   
                 365 
                 6364 
               
               
                   
                   
                   
                 367 
                 17545 
               
               
                   
                   
                   
                 369 
                 13124 
               
               
                   
                   
                   
                 371 
                 17587 
               
               
                   
                   
                   
                 373 
                 36290 
               
               
                   
                   
                   
                 375 
                 5196 
               
               
                   
                   
                   
                 377 
                 5474 
               
               
                   
                   
                   
                 379 
                 6894 
               
               
                   
                   
                   
                 381 
                 9644 
               
               
                   
                   
                   
                 383 
                 21983 
               
               
                   
                   
                   
                 450 
                 29856 
               
               
                   
                   
                   
                 452 
                 10349 
               
               
                   
                   
                   
                 454 
                 18100 
               
               
                   
                   
                   
                 503 
                 19014 
               
               
                   
                   
               
               
                   
                 *RF = Relative Fluorescence 
               
            
           
         
       
     
     
       
         
           
               
             
               
                 TABLE 5 
               
             
            
               
                   
               
               
                 Relative Fluorescence of Transfection of C2C12 Cells 
               
            
           
           
               
               
               
               
               
            
               
                   
                 SEQ ID NO 
                 RF* 
                 SEQ ID NO 
                 RF 
               
               
                   
                   
               
            
           
           
               
               
               
               
               
            
               
                   
                 DOTMA 
                 1289 
                 328 
                 5507 
               
               
                   
                 205 
                 2102 
                 332 
                 537 
               
               
                   
                 216 
                 1001 
                 335 
                 4689 
               
               
                   
                 218 
                 3706 
                 336 
                 9647 
               
               
                   
                 219 
                 1346 
                 338 
                 2290 
               
               
                   
                 220 
                 1470 
                 341 
                 2141 
               
               
                   
                 224 
                 1469 
                 342 
                 9496 
               
               
                   
                 226 
                 1664 
                 343 
                 3218 
               
               
                   
                 229 
                 1164 
                 344 
                 1678 
               
               
                   
                 230 
                 2606 
                 345 
                 6569 
               
               
                   
                 234 
                 681 
                 347 
                 4457 
               
               
                   
                 236 
                 958 
                 348 
                 8625 
               
               
                   
                 236 
                 2142 
                 350 
                 2631 
               
               
                   
                 237 
                 6491 
                 351 
                 3530 
               
               
                   
                 238 
                 1550 
                 352 
                 3324 
               
               
                   
                 239 
                 10620 
                 353 
                 4291 
               
               
                   
                 256 
                 3207 
                 354 
                 1912 
               
               
                   
                 268 
                 2568 
                 355 
                 1900 
               
               
                   
                 323 
                 7698 
                 357 
                 4506 
               
               
                   
                 326 
                 8553 
                 358 
                 2271 
               
               
                   
                 327 
                 2043 
                 359 
                 5693 
               
               
                   
                   
                   
                 360 
                 2090 
               
               
                   
                   
                   
                 361 
                 585 
               
               
                   
                   
                   
                 363 
                 1080 
               
               
                   
                   
                   
                 365 
                 5390 
               
               
                   
                   
                   
                 367 
                 16207 
               
               
                   
                   
                   
                 369 
                 3322 
               
               
                   
                   
                   
                 371 
                 983 
               
               
                   
                   
                   
                 373 
                 3037 
               
               
                   
                   
                   
                 375 
                 5633 
               
               
                   
                   
                   
                 377 
                 1710 
               
               
                   
                   
                   
                 379 
                 3189 
               
               
                   
                   
                   
                 381 
                 8834 
               
               
                   
                   
                   
                 383 
                 1876 
               
               
                   
                   
                   
                 450 
                 3795 
               
               
                   
                   
                   
                 452 
                 21798 
               
               
                   
                   
                   
                 454 
                 9280 
               
               
                   
                   
                   
                 501 
                 5913 
               
               
                   
                   
                   
                 502 
                 2589 
               
               
                   
                   
                   
                 503 
                 3973 
               
               
                   
                   
               
               
                   
                 *RF = Relative Fluorescence 
               
            
           
         
       
     
     
       
         
           
               
             
               
                 TABLE 6 
               
             
            
               
                   
               
               
                 Relative Fluorescence of Transfection of Human Fibroblast Cells 
               
            
           
           
               
               
               
               
               
            
               
                   
                 SEQ ID NO 
                 RF* 
                 SEQ ID NO 
                 RF 
               
               
                   
                   
               
            
           
           
               
               
               
               
               
            
               
                   
                 DOTMA 
                 3397 
                 328 
                 5641 
               
               
                   
                 205 
                 8550 
                 332 
                 5178 
               
               
                   
                 216 
                 3117 
                 335 
                 8534 
               
               
                   
                 218 
                 11230 
                 336 
                 10449 
               
               
                   
                 219 
                 7389 
                 338 
                 5625 
               
               
                   
                 220 
                 4516 
                 341 
                 2284 
               
               
                   
                 224 
                 5995 
                 342 
                 9525 
               
               
                   
                 226 
                 3431 
                 343 
                 4590 
               
               
                   
                 229 
                 6811 
                 344 
                 12704 
               
               
                   
                 230 
                 11771 
                 345 
                 2816 
               
               
                   
                 234 
                 4145 
                 347 
                 9255 
               
               
                   
                 236 
                 3630 
                 348 
                 11802 
               
               
                   
                 236 
                 3000 
                 349 
                 8810 
               
               
                   
                 237 
                 2184 
                 350 
                 7105 
               
               
                   
                 238 
                 3348 
                 351 
                 8061 
               
               
                   
                 239 
                 4084 
                 352 
                 3059 
               
               
                   
                 256 
                 2656 
                 353 
                 8653 
               
               
                   
                 268 
                 4568 
                 354 
                 4998 
               
               
                   
                 323 
                 4826 
                 355 
                 6821 
               
               
                   
                 326 
                 3070 
                 357 
                 7167 
               
               
                   
                 327 
                 4662 
                 358 
                 5665 
               
               
                   
                   
                   
                 359 
                 3206 
               
               
                   
                   
                   
                 360 
                 5347 
               
               
                   
                   
                   
                 361 
                 7014 
               
               
                   
                   
                   
                 363 
                 3008 
               
               
                   
                   
                   
                 365 
                 5219 
               
               
                   
                   
                   
                 367 
                 10036 
               
               
                   
                   
                   
                 369 
                 7590 
               
               
                   
                   
                   
                 371 
                 4584 
               
               
                   
                   
                   
                 373 
                 4872 
               
               
                   
                   
                   
                 375 
                 7305 
               
               
                   
                   
                   
                 377 
                 6638 
               
               
                   
                   
                   
                 379 
                 8520 
               
               
                   
                   
                   
                 381 
                 9953 
               
               
                   
                   
                   
                 383 
                 5896 
               
               
                   
                   
                   
                 450 
                 6996 
               
               
                   
                   
                   
                 452 
                 2779 
               
               
                   
                   
                   
                 454 
                 7568 
               
               
                   
                   
                   
                 501 
                 9473 
               
               
                   
                   
                   
                 503 
                 13085 
               
               
                   
                   
               
               
                   
                 *RF = Relative Fluorescence 
               
            
           
         
       
     
     
       
         
           
               
             
               
                 TABLE 7 
               
             
            
               
                   
               
               
                 Relative Fluorescence of Transfection of Jurkat Cells 
               
            
           
           
               
               
               
               
               
            
               
                   
                 SEQ ID NO 
                 RF* 
                 SEQ ID NO 
                 RF 
               
               
                   
                   
               
            
           
           
               
               
               
               
               
            
               
                   
                 DOTMA 
                 9084 
                 328 
                 4770 
               
               
                   
                 205 
                 2298 
                 332 
                 3052 
               
               
                   
                 216 
                 3570 
                 335 
                 4567 
               
               
                   
                 218 
                 21451 
                 336 
                 4813 
               
               
                   
                 219 
                 10762 
                 338 
                 4475 
               
               
                   
                 220 
                 6428 
                 341 
                 18042 
               
               
                   
                 224 
                 13170 
                 342 
                 12825 
               
               
                   
                 226 
                 12976 
                 343 
                 23499 
               
               
                   
                 229 
                 8079 
                 344 
                 10160 
               
               
                   
                 230 
                 10229 
                 345 
                 2462 
               
               
                   
                 234 
                 5441 
                 347 
                 17720 
               
               
                   
                 236 
                 11006 
                 348 
                 15585 
               
               
                   
                 236 
                 4593 
                 349 
                 23167 
               
               
                   
                 237 
                 1501 
                 350 
                 9184 
               
               
                   
                 238 
                 6278 
                 351 
                 9730 
               
               
                   
                 239 
                 2343 
                 352 
                 3150 
               
               
                   
                 256 
                 3322 
                 353 
                 11107 
               
               
                   
                 268 
                 3236 
                 354 
                 16705 
               
               
                   
                 323 
                 2900 
                 355 
                 8132 
               
               
                   
                 326 
                 3018 
                 357 
                 2256 
               
               
                   
                 327 
                 14546 
                 358 
                 18820 
               
               
                   
                   
                   
                 359 
                 1880 
               
               
                   
                   
                   
                 360 
                 16607 
               
               
                   
                   
                   
                 361 
                 7006 
               
               
                   
                   
                   
                 363 
                 2432 
               
               
                   
                   
                   
                 365 
                 5298 
               
               
                   
                   
                   
                 367 
                 4365 
               
               
                   
                   
                   
                 369 
                 11085 
               
               
                   
                   
                   
                 371 
                 6093 
               
               
                   
                   
                   
                 373 
                 12628 
               
               
                   
                   
                   
                 375 
                 4272 
               
               
                   
                   
                   
                 377 
                 6785 
               
               
                   
                   
                   
                 379 
                 7711 
               
               
                   
                   
                   
                 381 
                 10806 
               
               
                   
                   
                   
                 383 
                 14910 
               
               
                   
                   
                   
                 450 
                 8779 
               
               
                   
                   
                   
                 452 
                 2554 
               
               
                   
                   
                   
                 454 
                 8224 
               
               
                   
                   
                   
                 501 
                 12350 
               
               
                   
                   
                   
                 503 
                 9411 
               
               
                   
                   
               
               
                   
                 *RF = Relative Fluorescence 
               
            
           
         
       
     
     
       
         
           
               
             
               
                 TABLE 8 
               
             
            
               
                   
               
               
                 Visual Confirmation of Transfection of Cells 
               
            
           
           
               
               
            
               
                 Cell Line 
                 SEQ ID NOs 
               
               
                   
               
               
                 Rat Cortical Neurons 
                 220, 236, 238, 323, 327, 336, 338, 341, 
               
               
                   
                 343, 347, 348, 350, 351, 352, 354, 367, 
               
               
                   
                 369, 373, 375, 377, 454 
               
               
                 THP-1 
                 219, 229, 230, 239, 323, 328, 332, 341, 
               
               
                   
                 343, 350, 351, 357, 358, 375, 450, 454 
               
               
                 Human Skeletal Muscle 
                 218, 219, 230, 328, 336, 344, 350, 351, 
               
               
                   
                 353, 355, 365, 375 
               
               
                   
               
            
           
         
       
     
     
       
         
           
               
             
               
                 TABLE 9 
               
             
            
               
                   
               
               
                 Relative Fluorescence DOTMA/SEQ ID NO: 72/Peptide as Compared 
               
               
                 with Lipofectamine 2000 for the Transfection of HeLa Cells 
               
            
           
           
               
               
               
            
               
                   
                 RF* 
                   
               
            
           
           
               
               
               
               
               
            
               
                   
                 0.1 †   
                 0.2 †   
                 0.3 †   
                 0.4 †   
               
               
                   
                   
               
            
           
           
               
               
               
               
               
            
               
                 Transfection 
                   
                   
                   
                   
               
               
                 Agent 
               
               
                 Lipo #   
                 12694 
                 22154 
                 21542 
                 19286 
               
               
                 DOTMA/72 §   
                 10178 
                 11133 
                 8021 
                 7032 
               
               
                 SEQ ID NO ‡   
               
               
                 218 
                 12686 
                 13103 
                 10151 
                 10081 
               
               
                 219 
                 12135 
                 13841 
                 13531 
                 11712 
               
               
                 236 
                 5993 
                 8846 
                 8492 
                 7316 
               
               
                 237 
                 8986 
                 9753 
                 8015 
                 8288 
               
               
                 268 
                 10162 
                 9934 
                 10792 
                 9519 
               
               
                 323 
                 15537 
                 16620 
                 10839 
                 9244 
               
               
                 326 
                 6887 
                 9487 
                 10330 
                 9227 
               
               
                 328 
                 11037 
                 12229 
                 10801 
                 10951 
               
               
                 335 
                 12866 
                 12458 
                 11547 
                 9753 
               
               
                 336 
                 14442 
                 16702 
                 13434 
                 11334 
               
               
                 341 
                 6406 
                 7469 
                 8020 
                 7546 
               
               
                 342 
                 41299 
                 36040 
                 28841 
                 18351 
               
               
                 343 
                 6961 
                 8626 
                 8577 
                 8160 
               
               
                 344 
                 13315 
                 16387 
                 12564 
                 12511 
               
               
                 347 
                 23188 
                 21493 
                 16814 
                 14140 
               
               
                 348 
                 17413 
                 18615 
                 16164 
                 15054 
               
               
                 350 
                 24092 
                 23238 
                 19802 
                 22255 
               
               
                 351 
                 7678 
                 11499 
                 10575 
                 8791 
               
               
                 353 
                 36315 
                 44772 
                 36882 
                 25802 
               
               
                 355 
                 18362 
                 27990 
                 22367 
                 18289 
               
               
                 357 
                 7718 
                 11798 
                 9229 
                 9107 
               
               
                 358 
                 12331 
                 12683 
                 13686 
                 11097 
               
               
                 360 
                 9946 
                 11168 
                 10381 
                 8995 
               
               
                 365 
                 26620 
                 43775 
                 36054 
                 21540 
               
               
                 367 
                 10427 
                 21411 
                 23001 
                 17529 
               
               
                 369 
                 8737 
                 16706 
                 14880 
                 12645 
               
               
                 375 
                 22457 
                 30891 
                 34887 
                 29024 
               
               
                 377 
                 39115 
                 40072 
                 32914 
                 22457 
               
               
                 379 
                 50018 
                 47049 
                 39036 
                 26448 
               
               
                 381 
                 34187 
                 37463 
                 27007 
                 20088 
               
               
                 450 
                 11962 
                 17346 
                 12553 
                 10241 
               
               
                 452 
                 6276 
                 12671 
                 12047 
                 10918 
               
               
                 454 
                 34033 
                 37099 
                 27412 
                 16023 
               
               
                 501 
                 23571 
                 23993 
                 20410 
                 14749 
               
               
                 503 
                 12827 
                 20006 
                 17101 
                 16206 
               
               
                   
               
               
                 *RF = Relative Fluorescence 
               
               
                   † Volume of DOTMA/SEQ ID NO: 72/Peptide in μL in well of a 96 well plate 
               
               
                   # Lipofectamine 2000 
               
               
                   § DOTMA and SEQ ID NO: 72 with no additional peptide 
               
               
                   ‡ Peptide mixed with DOTMA/SEQ ID NO: 72 
               
            
           
         
       
     
     Example 2: Plant Movement Proteins 
     Cells were plated to so that on the day of transfection the cells were 70% confluent in 96 well tissue culture plates. A DOMTA/DOPE Lipid solution (1:1 molar ratio) at 2 mg/mL in water were mixed with an equal volume of plant virus movement protein derived peptides that had the nucleic acid binding moiety RRRRRRRRRRRR (SEQ ID NO:109) or KKKKKKKKKKKKKKKK (SEQ ID NO: 102) covalently linked to the N-terminus, or C-terminus of each peptide during peptide synthesis. Peptides were at 2 mg/mL in water. The DOTMA/DOPE and the plant virus movement peptide solution the was diluted 1 to 1 (v/v) in water. DOMTA/DOPE/PEPTIDE solutions were added to 0.1 mL of a plasmid DNA solution (EF 1Alpha eGFP plasmid) at 10 μg/mL in OptiMEM. 
     All solutions were at room temperature. A volume of 0.2 mL of a 10 μg/mL solution of DNA was aliquoted into each well of a non-tissue cultured treated plate. 1-6 μL of transfection reagents were added to DNA solutions respectively. The transfection reagent and DNA solution was mixed by pipetting up and down twice. Transfection complexes were formed for 10 minutes. After 10 minutes, 0.01 or 0.02 mL of the transfection complex was added to cells. HuVEC, HeLa, human adult keratinocytes, human primary adult fibroblast, and A549 cells all respectively received 0.01 mL of the transfection complex. Human skeletal muscle cells, mouse C2C12 cells, and Jurkat, cells received 0.02 mL of the transfection complex. Cells were incubated for 42 hours at 37° C. at 5% CO 2 . Plates were read on a fluorescent plate reader. Cells were also examined visually under a microscope to assess the extent of transfection (in terms of the percent of cells transfected) with a fluorescent microscope. Other modes of analysis, for example quantification with B-galactosidase or luciferase reports plasmids can also be used. If the plate reader did not show a sufficient signal to noise ratio, then plates were scored for cells transfected and those peptides that show increase over DOTMA alone were noted. 
     
       
         
           
               
             
               
                 TABLE 10 
               
             
            
               
                   
               
               
                 Relative Fluorescence of Transfection of Hela Cells 
               
            
           
           
               
               
            
               
                 Transfection 
                   
               
               
                 Agent 
               
               
                 SEQ ID NO 
               
               
                   
               
            
           
           
               
               
               
               
               
               
               
            
               
                 543 
                 12174 
                 42502 
                 38779 
                 28451 
                 13793 
                 7705 
               
               
                 545 
                 11056 
                 28529 
                 28943 
                 14693 
                 13009 
                 10902 
               
               
                 544 
                 10683 
                 19151 
                 19877 
                 16908 
                 12759 
                 8938 
               
               
                 540 
                 6422 
                 35653 
                 27970 
                 18620 
                 13917 
                 10336 
               
               
                 541 
                 17145 
                 36476 
                 37105 
                 23644 
                 20473 
                 15523 
               
               
                 542 
                 9010 
                 32901 
                 47293 
                 42152 
                 35631 
                 18856 
               
               
                 DOTMA 
                 4424 
                 4782 
                 5377 
                 6992 
                 9694 
                 11720 
               
               
                 1 μg/ml 
               
               
                 109 
                 4157 
                 5578 
                 6606 
                 8520 
                 8007 
                 7803 
               
               
                 547 
                 5899 
                 9491 
                 10275 
                 11241 
                 8829 
                 7989 
               
               
                 546 
                 15190 
                 15022 
                 12772 
                 11535 
                 8117 
                 7181 
               
               
                 Blank 
                 3912 
                 3941 
                 3860 
                 3805 
                 3990 
                 3871 
               
               
                   
               
            
           
         
       
     
     
       
         
           
               
             
               
                 TABLE 11 
               
             
            
               
                   
               
               
                 Relative Fluorescence of Transfection 
               
               
                 of Human Primary Fibroblast Cells 
               
            
           
           
               
               
            
               
                   
                 Transfection Agent 
               
               
                   
                 Rf 
               
            
           
           
               
               
               
               
               
               
               
            
               
                 SEQ ID NO 
                 1 μl 
                 1.5 μl 
                 2 μl 
                 3 μl 
                 4 μl 
                 6 μl 
               
               
                   
               
            
           
           
               
               
               
               
               
               
               
            
               
                 Blank 
                 26467 
                 26415 
                 26510 
                 26317 
                 26738 
                 26620 
               
               
                 543 
                 30416 
                 45401 
                 39786 
                 33091 
                 28648 
                 28698 
               
               
                 545 
                 34309 
                 48184 
                 37497 
                 29067 
                 28928 
                 29256 
               
               
                 544 
                 31368 
                 39311 
                 37634 
                 32892 
                 31428 
                 28367 
               
               
                 540 
                 29368 
                 40610 
                 35505 
                 33853 
                 33063 
                 30219 
               
               
                 541 
                 35030 
                 44894 
                 42097 
                 31837 
                 29537 
                 28921 
               
               
                 542 
                 30818 
                 43609 
                 46966 
                 36343 
                 33706 
                 31787 
               
               
                 DOTMA 
                 28527 
                 28021 
                 28035 
                 28646 
                 29613 
                 34911 
               
               
                 1 μg/ml 
               
               
                 109 
                 28368 
                 30373 
                 29858 
                 29895 
                 30941 
                 29386 
               
               
                 547 
                 29141 
                 33704 
                 35175 
                 35563 
                 33951 
                 30539 
               
               
                 546 
                 31980 
                 39259 
                 41747 
                 35020 
                 32220 
                 29476 
               
               
                 Blank 
                 26822 
                 26658 
                 26624 
                 25988 
                 27112 
                 26703 
               
               
                   
               
            
           
         
       
     
     
       
         
           
               
             
               
                 TABLE 12 
               
             
            
               
                   
               
               
                 Relative Fluorescence of Transfection of HuVEC Cells 
               
            
           
           
               
               
            
               
                   
                 Transfection Agent 
               
               
                   
                 Rf 
               
            
           
           
               
               
               
               
               
               
               
            
               
                 SEQ ID NO 
                 1 μl 
                 1.5 μl 
                 2 μl 
                 3 μl 
                 4 μl 
                 6 μl 
               
               
                   
               
            
           
           
               
               
               
               
               
               
               
            
               
                 Blank 
                 23420 
                 23592 
                 23275 
                 23437 
                 23969 
                 23973 
               
               
                 543 
                 26622 
                 36742 
                 42802 
                 40584 
                 32754 
                 25492 
               
               
                 545 
                 26545 
                 36649 
                 46890 
                 34047 
                 30893 
                 27133 
               
               
                 544 
                 25788 
                 28622 
                 31245 
                 34718 
                 31159 
                 28639 
               
               
                 540 
                 24771 
                 34062 
                 44744 
                 44087 
                 45137 
                 30023 
               
               
                 541 
                 27405 
                 46446 
                 OVER 
                 40401 
                 31568 
                 28125 
               
               
                 542 
                 24827 
                 33896 
                 42546 
                 43231 
                 35294 
                 29326 
               
               
                 DOTMA 
                 24590 
                 24571 
                 24811 
                 24463 
                 26972 
                 29850 
               
               
                 1 μg/ml 
               
               
                 109 
                 24603 
                 25735 
                 25421 
                 26641 
                 26529 
                 25907 
               
               
                 547 
                 24554 
                 26172 
                 28255 
                 32494 
                 31180 
                 26775 
               
               
                 546 
                 25990 
                 32762 
                 37899 
                 35528 
                 31614 
                 27690 
               
               
                 Blank 
                 24151 
                 23427 
                 23055 
                 23320 
                 23689 
                 24147 
               
               
                   
               
            
           
         
       
     
     Example 3: Formulation 1 
     Formulation 1 comprised a combination of cationic lipids, neutral lipids, and peptides, as described above. Specifically, the cationic lipids used in Formulation 1 included DPePE-Sp-OMe and DPB-Arg. These cationic lipids are compounds of Formula 1, where:
         DPePE-Sp-OMe=R 1 ═R 2 ═C 16 H 31 , X═O, W 1 ═H; W 2 ═CH 2 —O—P(═O)(OMe)-O—CH 2 CH 2 —NH—C(═O)-spermine, y=0;   DPB-Arg=W 1 ═W 2 ═CH 2 —N(R 3 R 4 ), R 1 ═R 2 ═C 16 H31, X═O; R 3 ═CH 3 , R 4 ═—C(═O)—C(NH 2 )—CH 2 —(CH 2 ) n —C(═NH)—NH 2 , n=0-6, y=0.       

     The neutral lipids included dioleoylphosphatidylethanolamine (DOPE), diphytanoylphosphatidylethanolamine (DPhPE), and 3-oleyloxy-2-hydroxy-1-acetamidopropane (HOAP). 
     The cationic lipid and the neutral lipid are combined in ethanol in the ratio of 1:8 (cationic lipid:neutral lipid). Formulation 1 was obtained by mixing the lipid with water and the peptide in the ratio of 0.2:1:1 (lipid mix:water:peptide mix). Specifically, the following peptides were used to prepare Formulation 1: 
     Peptide A: SEQ ID NO:507 covalently linked to SEQ ID NO:109 (4.1 mg/mL in HEPES);
 
Peptide B: SEQ ID NO:540 (4.1 mg/mL in HEPES);
 
Peptide C: SEQ ID NO:350 covalently linked to SEQ ID NO:109 (4.1 mg/mL in HEPES);
 
Peptide D: SEQ ID NO:47 (1 mg/mL in TRIS).
 
     For Formulation 1, the lipid component comprised lipids in the molar ratio of 0.50:0.50:2.666:2.666:2.666 (DPePe-Sp-OMe:DPB-Arg: DOPE:DPhPE:HOAP). This translates to a cationic lipid:neutral lipid ratio of 1:8. The mixture of peptides in the formulation were present in the ratio of 1:2:2:0.5 by volume (Peptide A:Peptide B:Peptide C:Peptide D). 
     Example 4: Transfection 
     Human Adult Dermal Fibroblast was obtained from ScienCell Research Laboratories (Carlsbad, Calif.) and grown according to manufacturer&#39;s instructions. Human Adult Keratinocytes were obtained from ThermoFisher Scientific and grown according to manufactures instructions in Keratinocyte-SFM. Cells were plated to obtain approximately 70% confluency the day of transfection in 96-well tissue culture treated plates. Green Fluorescent Protein expression plasmid was used in these experiments. Transfection complexes for Lipofectamine 3000 were prepared according to manufacturer&#39;s instructions. All solutions used to form transfection complexes were at room temperature for transfection. Into each well of a non-tissue culture treated plate containing 0.05 mL of OptIMEM were aliquoted 1.0, 1.5, 2.0, 3.0, 4.0 and 6.0 μL of the Formulation 1 transfection reagent. A volume of 0.05 mL of a 20 μg/mL solution of DNA in OptiMEM was aliquoted into each well of a non-tissue culture treated plate that contained transfection reagent aliquots. The transfection reagent and DNA solution were mixed by pipetting up and down twice. Transfection complexes were formed for 10 minutes at room temperature. After 10 minutes, 0.01 mL of the transfection complex was added to cells. Cells were incubated for 42 hours at 37° C. at 5% CO 2 . Plates were read on a fluorescent plate reader. The results are shown in Tables 13 and 14. Cells were also examined visually under a microscope to assess the extent of transfection (regarding the percent of cells transfected) with a fluorescent microscope. Other modes of analysis, for example, quantification with B-galactosidase or luciferase reports plasmids, can also be used. 
     
       
         
           
               
             
               
                 TABLE 13 
               
             
            
               
                   
               
               
                 Relative Fluorescence of Transfection 
               
               
                 of Human Primary Adult Fibroblasts 
               
            
           
           
               
               
               
               
               
            
               
                   
                 Volume of 
                   
                   
                   
               
               
                   
                 Transfection 
               
               
                   
                 Reagent/well 
                 Formulation 1 
                 Lipofectamine 3000 
                 Blank 
               
               
                   
                   
               
            
           
           
               
               
               
               
               
            
               
                   
                 0.1 μL 
                 21851 
                 9648 
                 9373 
               
               
                   
                 0.15 μL  
                 21359 
                 10041 
                 9017 
               
               
                   
                 0.2 μL 
                 26793 
                 12509 
                 9135 
               
               
                   
                 0.3 μL 
                 35137 
                 15193 
                 9088 
               
               
                   
                 0.4 μL 
                 42224 
                 16971 
                 9200 
               
               
                   
                 0.6 μL 
                 42154 
                 18095 
                 9345 
               
               
                   
                   
               
            
           
         
       
     
     
       
         
           
               
             
               
                 TABLE 14 
               
             
            
               
                   
               
               
                 Relative Fluorescence of Transfection 
               
               
                 of Human Primary Adult Keratinocytes 
               
            
           
           
               
               
               
               
            
               
                 Volume of 
                   
                   
                   
               
               
                 Transfection 
               
               
                 Reagent/well 
                 Formulation 1 
                 Lipofectamine 3000 
                 Blank 
               
               
                   
               
               
                 0.1 μL 
                 41003 
                 15028 
                 13843 
               
               
                 0.15 μL  
                 36375 
                 15981 
                 13820 
               
               
                 0.2 μL 
                 33300 
                 15538 
                 13674 
               
               
                 0.3 μL 
                 23132 
                 14780 
                 13412 
               
               
                 0.4 μL 
                 19578 
                 15176 
                 13987 
               
               
                 0.6 μL 
                 17126 
                 14934 
                 13704