Patent Publication Number: US-2022235374-A1

Title: Gene therapy for alzheimer`s disease

Description:
CLAIM OF PRIORITY 
     This application claims the benefit of U.S. Provisional Patent Application Ser. No. 62/852,716, filed on May 24, 2019. The entire contents of the foregoing are hereby incorporated by reference. 
    
    
     FEDERALLY SPONSORED RESEARCH OR DEVELOPMENT 
     This invention was made with Government support under Grant No. NS041783 awarded by the National Institutes of Health. The Government has certain rights to the invention. 
    
    
     TECHNICAL FIELD 
     Described herein are, inter alia, compositions and methods for using presenilin genetic therapy constructs to treat Alzheimer&#39;s disease (AD) and other neurodegenerative diseases. 
     BACKGROUND 
     Alzheimer&#39;s disease, also known as Alzheimer disease, accounts for majority of neurodegenerative dementia and is the fourth leading cause of death in the United States after heart disease, cancer and stroke. It is characterized by a progressive loss of cognitive function, neurodegeneration, neurofibrillary tangles and amyloid plaques in the brains of patients. Although the progression speed varies in different patients, the average life expectancy following diagnosis is three to nine years. Currently, there is no treatment for Alzheimer&#39;s disease. 
     SUMMARY 
     Described herein are methods and compositions that can be used to treat subjects with Alzheimer&#39;s disease (AD) and other neurodegenerative diseases, disorders or conditions. The present disclosure is based, at least in part, on the discovery that providing a codon-optimized wild-type PSEN1 cDNA into cells carrying heterozygous or homozygous dominant negative Psenl mutations, a well-established familial Alzheimer&#39;s disease model, provided unexpectedly high expression levels and rescued the impaired γ-secretase activity in these cells. Thus, the present disclosure provides methods for effective gene therapy based on PSEN1 (to express PS1) and/or PSEN2 (to express PS2) for Alzheimer&#39;s disease and other neurodegenerative dementia, representing a significant breakthrough in this disease area. 
     Provided herein are compositions comprising a human codon-optimized polynucleotide encoding a human presenilin 1 protein (PS1), e.g., a polynucleotide comprising SEQ ID NO:9 or a sequence that is at least 80%, 90%, 95%, or 99% identical to SEQ ID NO:9 (with at least one codon optimized with respect to wild type). Exemplary human PS1 protein sequences include SEQ ID NOs:5 and 6, and can include sequences that are at least comprising a human codon-optimized polynucleotide encoding a human presenilin 1 (PSEN1) thereto. In some embodiments, the composition is associated with (e.g., formulated for delivery using) an exosome or lipid-based nanoparticle (LNP). 
     Also provided herein are compositions comprising a vector for expression of human PSEN1 in a cell, comprising a human codon-optimized polynucleotide described herein, operably linked to a promoter. 
     Also provided herein is the use of any of the compositions described herein in a method o treating a neurodegenerative disease, disorder, or condition in a subject. 
     In some embodiments, the vector is a viral vector, e.g., an adeno-associated viral (AAV) vector (such as AAV9 or AAVrh10); a lentiviral vector; or a retroviral vector. 
     In some embodiments, the promoter is a pan neuronal promoter, e.g., a synapsin I promoter, or a neuron subtype-specific promoter, e.g., an alpha-calcium/calmodulin kinase 2A promoter. 
     Also provided herein are methods for treating a neurodegenerative disease, disorder, or condition, the method comprising administering to a human subject in need of treatment a composition described herein, wherein the subject has one or more mutations in at least one allele of PSEN1, preferably a mutation that encodes a dominant negative PSEN1 protein isoform. 
     In some embodiments, the neurodegenerative disease, disorder or condition is Alzheimer&#39;s disease. 
     In some embodiments, the Alzheimer&#39;s disease is familial Alzheimer&#39;s disease. In some embodiments, the Alzheimer&#39;s disease is late-onset Alzheimer&#39;s disease. In some embodiments, the Alzheimer&#39;s disease is sporadic Alzheimer&#39;s disease. In some embodiments, the Alzheimer&#39;s disease is early-onset Alzheimer&#39;s disease. 
     In some embodiments, the subject has a mutation at E280, Y115, L166, C410, Δex9, G548, D257, R278, L435, G384, L392, N141, G206, H163, A79, S290, A260, A426, A431, R269, L271, C1410, E280, P264, E185, L235, M146, e.g., a E280A, Y115H, L166P, C410Y, Δex9, G548, D257A, R278I, L435F, G384A, or L392V mutation in the PSEN1 gene, or a N141I, G206A, H163R, A79V, S290C, A260P, A426P, A431E, R269H, L271V, C1410Y, E280G, P264L, E185D, L235V, or M146V mutation in the PSEN1 gene. 
     In some embodiments, the neurodegenerative disease, disorder or condition is frontotemporal dementia, memory loss, cognitive decline or impairment. In some embodiments, the cognitive impairment is mild cognitive impairment (MCI). 
     In some embodiments, the composition is administered to the CNS of the subject in need of treatment. 
     In some embodiments, the polynucleotide encoding PSEN1 and/or PSEN2 gene or mRNA is administered to the CNS via intravenous delivery, via intrathecal delivery, via intracisternal delivery, via intracerebroventricular delivery, or via stereotactic injection into certain areas of the brain, optionally into the cerebral ventricles, or via direct injection into hippocampus or cortical areas 
     Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs. Methods and materials are described herein for use in the present invention; other, suitable methods and materials known in the art can also be used. The materials, methods, and examples are illustrative only and not intended to be limiting. All publications, patent applications, patents, sequences, database entries, and other references mentioned herein are incorporated by reference in their entirety. In case of conflict, the present specification, including definitions, will control. 
     Other features and advantages of the invention will be apparent from the following detailed description and figures, and from the claims. 
    
    
     
       DESCRIPTION OF DRAWINGS 
         FIGS. 1A-B . Introduction of WT hPS1 rescues impaired γ-secretase activity in mutant MEFs. A, γ-Secretase activity measured by NICD production is reduced in mutant MEF cells in a PS dosage dependent manner (WT&gt;PS1 heterozygous KI or KO&gt;homozygous PS1 KI or KO&gt;DKO). B, Restoring impaired γ-secretase activity by WT hPS1. Increasing amounts of pCI-hPSEN1 plasmid DNA, as indicated, are transfected into MEFs of varying genotypes. Western analysis showed that both PS1 NTF and NICD are restored in various PS mutant MEFs. Heterozygous L435F KI cells are labeled as KI/+ or PS1L435F/+. N=3 independent experiments. Data represent mean±SEM. *p&lt;0.05; **p&lt;0.01; ***p&lt;0.001 (one-way ANOVA with Tukey&#39;s post-hoc analysis). 
         FIG. 2 . Sequence comparison of the endogenous human PSEN1 (hPSEN1) cDNA and the codon optimized hPSEN1 cDNA (Opti-hPSEN1) 
         FIGS. 3A-B . Increased expression levels of PS1 NTF with codon optimized PSEN1 cDNA. A, Psen-null MEFs were transfected with increasing amounts of plasmids expressingexpressing the wild-type endogenous hPSEN1 cDNA (wt_PS1) or the codon optimized hPSEN1 cDNA (opti_PS1), and Western analysis was carried out using an antibody specific for PS1 N terminus. Untrans: untransfected MEFs as negative control. B, Quantification of PS1 NTF levels in cells transfected with either wild-type endogenous hPSEN1 cDNA (wt_PS1) or the codon optimized hPSEN1 cDNA (opti_PS1). Data are expressed as Mean±SEM (n=3 independent experiments) 
         FIG. 4 . Codon optimization led to increased γ-secretase activity. PS DKO MEFs were transfected with increasing amounts (12.5, 25, 50 or 100 ng) of pCI-hPS1 or pCI-hPS1opti plasmid DNA and CMV-NΔE followed by Western analysis of NICD. MEFs that were untransfected or transfected with the empty vector were included as negative controls. We found that pCI-hPS1opti led to significantly higher levels of γ-secretase activity, measured by NICD production, relative to pCI-hPS1. Data are expressed as mean±SEM (n=5 independent experiments). Two-way ANOVA was used to assess statistical significance. **p&lt;0.01. 
     
    
    
     DEFINITIONS 
     In order for the present disclosure to be more readily understood, certain terms are first defined below. Additional definitions for the following terms and other terms are set forth throughout the specification. 
     Administration: 
     As used herein, the term “administration” refers to the delivery or application of a composition to a subject or system. Administration to an animal subject (e.g., to a human) may be by any appropriate route. For example, in some embodiments, administration may be bronchial (including by bronchial instillation), buccal, enteral, intra-arterial, intradermal, intragastric, intramedullary, intramuscular, intranasal, intraperitoneal, intrathecal, intravenous, intraventricular, mucosal, nasal, oral, rectal, subcutaneous, sublingual, topical, tracheal (including by intratracheal instillation), transdermal, vaginal and vitreal. 
     Biologically Active: 
     As used herein, the phrase “biologically active” refers to a characteristic of any substance that has activity in a biological system (e.g., cell culture, organism, etc.). For instance, a substance that, when administered to an organism, has a biological effect on that organism, is considered to be biologically active. Biological activity can also be determined by in vitro assays (for example, in vitro enzymatic assays). In particular embodiments, where a protein or polypeptide is biologically active, a portion of that protein or polypeptide that shares at least one biological activity of the protein or polypeptide is typically referred to as a “biologically active” portion. In some embodiments, a protein is produced and/or purified from a cell culture system, which displays biologically activity when administered to a subject. 
     Control: 
     As used herein, the term “control” has its art-understood meaning of being a standard against which results are compared. Typically, controls are used to augment integrity in experiments by isolating variables in order to make a conclusion about such variables. In some embodiments, a control is a reaction or assay that is performed simultaneously with a test reaction or assay to provide a comparator. In one experiment, the “test” (i.e., the variable being tested) is applied. In the second experiment, the “control,” the variable being tested is not applied. In some embodiments, a control is a historical control (i.e., of a test or assay performed previously, or an amount or result that is previously known). In some embodiments, a control is or comprises a printed or otherwise saved record. A control may be a positive control or a negative control. In some embodiments, the control may be a “reference control”, which is a sample used for comparison with a test sample, to look for differences or for the purposes of characterization. 
     Gene Therapy: 
     As used herein, the term “gene therapy” refers to any treatment including the direct or indirect administration of a nucleic acid to a subject. In particular instances, a protein of therapeutic value is expressed from an administered nucleic acid. 
     Identity: 
     As used herein, the term “identity” refers to the overall relatedness between polymeric molecules, e.g., between nucleic acid molecules (e.g., DNA molecules and/or RNA molecules) and/or between polypeptide molecules. Calculation of the percent identity of two nucleic acid sequences, for example, can be performed by aligning the two sequences for optimal comparison purposes (e.g., gaps can be introduced in one or both of a first and a second nucleic acid sequences for optimal alignment and non-identical sequences can be disregarded for comparison purposes). In certain embodiments, the length of a sequence aligned for comparison purposes is at least 30%, at least 40%, at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, at least 95%, or substantially 100% of the length of the reference sequence. The nucleotides at corresponding nucleotide positions are then compared. When a position in the first sequence is occupied by the same nucleotide as the corresponding position in the second sequence, then the molecules are identical at that position. The percent identity between the two sequences is a function of the number of identical positions shared by the sequences, taking into account the number of gaps, and the length of each gap, which needs to be introduced for optimal alignment of the two sequences. The comparison of sequences and determination of percent identity between two sequences can be accomplished using a mathematical algorithm. For example, the percent identity between two nucleotide sequences can be determined using the algorithm of Meyers and Miller (CABIOS, 1989, 4: 11-17), which has been incorporated into the ALIGN program (version 2.0) using a PAM120 weight residue table, a gap length penalty of 12 and a gap penalty of 4. The percent identity between two nucleotide sequences can, alternatively, be determined using the GAP program in the GCG software package using an NWSgapdna.CMP matrix. Various other sequence alignment programs are available and can be used to determine sequence identity such as, for example, Clustal. 
     Improve, Increase, or Reduce: 
     As used herein, the terms “improve,” “increase” or “reduce,” or grammatical equivalents, indicate values that are relative to a baseline measurement, such as a measurement in the same individual prior to initiation of the treatment described herein, or a measurement in a control individual (or multiple control individuals) in the absence of the treatment described herein. A “control individual” is an individual afflicted with the same type and approximately the same severity of, e.g., Alzheimer&#39;s disease, as the individual being treated, who is about the same age as the individual being treated (to ensure that the stages of the disease in the treated individual and the control individual(s) are comparable). 
     Neurodegeneration: 
     As used herein, the term “neurodegeneration” means a process in which one or more neurons are damaged, decrease in function, become dysfunctional, and/or are lost by cell death. Neurodegeneration encompasses both rapid, gradual, and intermediate forms. Accordingly, a neurodegenerative disease, condition, or symptom is one characterized in that the disease is typically associated with neuronal damage, and/or cell death. 
     Subject: 
     As used herein, the term “subject” refers to a human or any non-human animal (e.g., a mammal such as a mouse, rat, rabbit, dog, cat, cattle, swine, sheep, horse or primate). A human includes pre- and post-natal forms. In many embodiments, a subject is a human being. A subject can be a patient, which refers to a human presenting to a medical provider for diagnosis or treatment of a disease. The term “subject” is used herein interchangeably with “individual” or “patient.” A subject can be afflicted with or is susceptible to a disease or disorder but may or may not display symptoms of the disease or disorder. 
     Suffering from: 
     An individual who is “suffering from” a disease, disorder, and/or condition (e.g., Alzheimer&#39;s disease) has been diagnosed with or displays one or more symptoms of the disease, disorder, and/or condition. 
     Susceptible to: 
     An individual who is “susceptible to” a disease, disorder, and/or condition has not been diagnosed with and/or may not exhibit symptoms of the disease, disorder, and/or condition. In some embodiments, an individual who is susceptible to a disease, disorder, and/or condition (for example, Alzheimer&#39;s disease) may be characterized by one or more of the following: (1) a genetic mutation associated with development of the disease, disorder, and/or condition; (2) a genetic polymorphism associated with development of the disease, disorder, and/or condition; (3) increased and/or decreased expression and/or activity of a protein associated with the disease, disorder, and/or condition; (4) habits and/or lifestyles associated with development of the disease, disorder, and/or condition; (5) a family history of the disease, disorder, and/or condition; (6) reaction to certain bacteria or viruses; (7) exposure to certain chemicals. In some embodiments, an individual who is susceptible to a disease, disorder, and/or condition will develop the disease, disorder, and/or condition. In some embodiments, an individual who is susceptible to a disease, disorder, and/or condition will not develop the disease, disorder, and/or condition. 
     Therapeutically Effective Amount: 
     As used herein, the term “therapeutically effective amount” refers to an amount of a therapeutic protein which confers a therapeutic effect on the treated subject, at a reasonable benefit/risk ratio applicable to any medical treatment. The therapeutic effect may be objective (i.e., measurable by some test or marker) or subjective (i.e., subject gives an indication of or feels an effect). In particular, the “therapeutically effective amount” refers to an amount of a therapeutic protein or composition effective to treat, ameliorate, or prevent a desired disease or condition, or to exhibit a detectable therapeutic or preventative effect, such as by ameliorating symptoms associated with the disease, preventing or delaying the onset of the disease, and/or also lessening the severity or frequency of symptoms of the disease. A therapeutically effective amount is commonly administered in a dosing regimen that may comprise multiple unit doses. For any particular therapeutic protein, a therapeutically effective amount (and/or an appropriate unit dose within an effective dosing regimen) may vary, for example, depending on route of administration, on combination with other pharmaceutical agents. Also, the specific therapeutically effective amount (and/or unit dose) for any particular patient may depend upon a variety of factors including the disorder being treated and the severity of the disorder; the activity of the specific pharmaceutical agent employed; the specific composition employed; the age, body weight, general health, sex and diet of the patient; the time of administration, route of administration, and/or rate of excretion or metabolism of the specific fusion protein employed; the duration of the treatment; and like factors as is well known in the medical arts. 
     Treatment: 
     As used herein, the term “treatment” (also “treat” or “treating”), in its broadest sense, refers to any administration of a substance (e.g., provided compositions) that partially or completely alleviates, ameliorates, relives, inhibits, delays onset of, reduces severity of, and/or reduces incidence of one or more symptoms, features, and/or causes of a particular disease, disorder, and/or condition. In some embodiments, such treatment may be administered to a subject who does not exhibit signs of the relevant disease, disorder and/or condition and/or of a subject who exhibits only early signs of the disease, disorder, and/or condition. Alternatively or additionally, in some embodiments, treatment may be administered to a subject who exhibits one or more established signs of the relevant disease, disorder and/or condition. In some embodiments, treatment may be of a subject who has been diagnosed as suffering from the relevant disease, disorder, and/or condition. In some embodiments, treatment may be of a subject known to have one or more susceptibility factors that are statistically correlated with increased risk of development of the relevant disease, disorder, and/or condition. 
     Although generally speaking “PS1” refers to the presenilin-1 protein, and “PS2” refers to the presenilin-2 protein, in some cases PS1 or PS2 is used to refer to mRNA or gene. 
     DETAILED DESCRIPTION 
     The present disclosure provides, among other things, compositions and methods for treating subjects with Alzheimer&#39;s disease and other neurodegenerative diseases, disorders and conditions based on delivering functional presenilin-1 (PS1) protein to a subject in need thereof. In particular, the present disclosure contemplates gene therapy by providing a human codon-optimized polynucleotide encoding a presenilin-1 (PS1) to a subject in need of treatment who has a PSEN1 or PSEN2 mutation, e.g., a dominant negative mutation, associated with AD, e.g., with early onset Familial Alzheimer&#39;s disease (FAD) or with late onset sporadic AD. 
     Various aspects of the invention are described in detail in the following sections. The use of sections is not meant to limit the invention. Each section can apply to any aspect of the invention. In this application, the use of “or” means “and/or” unless stated otherwise. 
     Methods of Treatment 
     As non-limiting examples, the present methods include gene therapy to express wild-type human Presenilin-1 in a subject suffering from or susceptible to a neurodegenerative disease, e.g., associated with a dominant negative mutation in PSEN1 or PSEN2, e.g., Alzheimer&#39;s disease (e.g., familial AD patients carrying PSEN1 or PSEN2 mutations or sporadic AD patients). The objective of such a gene therapy is, among other things, to enhance expression of PS1 in the brains of familial or sporadic AD patients in order to correct or overcome a deficit in PS1 or PS2 expression and/or activity. In FAD patients, it is expected that a gene therapy method described herein results in increased expression of wild-type PS1 in the brain, rescuing the impairment of γ-secretase activity associated with PS1 or PS2 mutations. 
     Mutations in the Presenilin genes—PSEN1 and PSEN2—are highly penetrant and account for ˜90% of all mutations identified in familial AD (FAD), highlighting their importance in the pathogenesis of AD. More than 260 distinct mutations in PSEN1 have been reported, and they are dominantly inherited and mostly missense mutations. It is known that dominant negative mutations in the PSEN1 and PSEN2 genes are associated with early onset familial Alzheimer&#39;s disease. It was generally believed that the PS1 and presenilin-2 (PS2) proteins are part of □-secretase complex, and that mutations in the PSEN1 and PSEN2 genes contribute to the accumulation of Amyloid beta (Aβ) protein in Alzheimer&#39;s disease patients. Pathogenic PSEN1 mutations act in cis to impair mutant PS1 function and act in trans to inhibit wild-type Presently-1 (PS1) function (Heilig et al. J Neurosci 33:11606-717 (2013); Zhou et al. Proc Natl Acad Sci USA 114:12731-12736 (2017). Typically, by their very nature, dominant negative mutations cannot be rescued by expression of wild type protein (Herskowitz, I. Nature, 329:219-222 (1987)). However, surprisingly, as shown herein, transfection of codon-optimized hPSEN1 cDNA into immortalized MEFs carrying heterozygous and homozygous PS1 mutations was able to rescue the impaired γ-secretase activity in these cells even better than the wild-type human sequence (see Examples, below), indicating that expression of wild-type PS1 from a codon-optimized exogenous sequence can overcome the dominant negative effects of the mutant Presenilin protein. Without wishing to be bound by any particular theory, expression of PS1 may accomplish this objective by increasing the total level of wild-type PS1, rescuing the impairment of γ-secretase expression and/or activity in AD patients. 
     The methods and compositions described herein can equally be used to treat other neurodegenerative diseases, disorders or conditions. 
     Alzheimer&#39;s Disease 
     The methods described herein may be used to treat or reduce the risk of developing subjects with all types of Alzheimer&#39;s disease including, but not limited to, familial and sporadic Alzheimer&#39;s disease, early onset or late onset Alzheimer&#39;s disease. In some embodiments, the present methods may be used to treat or reduce the risk of development of early onset familial form of Alzheimer&#39;s disease (AD) that is associated with mutations in presenilin-1 (PS1) and/or presenilin-2 (PS2) (Sherrington, et al., Nature 375:754-760 (1995); Rogaev, et al., Nature 376:775-778 (1995); Levy-Lahad, et al., Science 269:970-973 (1995); Hiltunen, et al., Eur. J. Hum. Genet. 8:259-266 (2000); Jonghe, et al., Hum. Mol. Genet. 8:1529-1540 (1999); Tysoe, et al., Am. J. Hum. Genet. 62:70-76 (1998); Crook, et al., Nat. Med. 4:452-455 (1998), all of which are incorporated by reference herein). 
     In some embodiments, the present methods may be used to treat a subject that has a mutation in the PSEN1 or PSEN2 allele, e.g., a mutation that has a dominant negative effect on wild-type PS proteins. Exemplary mutations include C410Y, Δex9, G548, D257A, L166P, R278I, L435F, G384A, Y115H, and L392V, as well as N141I, G206A, H163R, A79V, S290C, A260P, A426P, A431E, R269H, L271V, C1410Y, E280G, P264L, E185D, L235V, and M146V mutations (see, e.g., Heilig et al., J. Neurosci., 33(28):11606-11617 (2013); Watanabe et al., J. Neurosci. 32(15):5085-5096 (2012); Brouwers et al., 2008 Ann Med 40 (8): 562-83); Watanabe and Shen, PNAS Nov. 28, 2017 114 (48) 12635-12637; Zhou et al., PNAS Nov. 28, 2017 114 (48) 12731-12736; Hsu et al., Alzheimers Res Ther. 2018 Jul. 18; 10(1):67). Additional exemplary mutations that may have a dominant negative effect on wild-type PS proteins can include, but are not limited to, in PSEN-1: N32N; R35Q; D40del (delGAC); D40del (delACG); E69D; A79V; V82L; I83_M84del (DelIM, ΔI83/M84, ΔI83/ΔM84); I83T; M84V; L85P; P88L; V89L (G&gt;T); V89L (G&gt;C); C92S; V94M; V96F; V97L; T99A; F105C; F105I; F105L; F105V; R108Q; L113_I114insT (Intron4, InsTAC, p.113+1delG, splice5); L113P; L113Q; Y115C; Y115D; Y115H; T116I; T116N; T116R; P117A; P117L; P117R; P117S; E120D (A&gt;C); E120D (A&gt;T); E120G; E120K; E123K; Q127_R128del(CAGA); InsG(G) (c.379_382del XXXXinsG); H131R; S132A; L134R; N135D; N135S; N135Y; A136G; M139I (G&gt;C); M139I (G&gt;A); M139K; M139L; M139T; M139V; V142F; I143F; I143M; I143N; I143T; I143V; M146I (G&gt;C); M146I (G&gt;T); M146I (G&gt;A); M146L (A&gt;C); M146L (A&gt;T); M146V; T147I; T147P; L150P; L153V; Y154C; Y154N; Y156F; Y156_R157insIY; R1575; H163P; H163R; H163Y; A164V; W165C (G&gt;C); W165C (G&gt;T); W165G; L166H; L166P; L166R; L166V; L166del; I167del (TTAdel); I167del (TATdel); I168T; S169del (ΔS169, Ser169del, ΔS170); S169L; S169P; S170F; S170P; L171P; L173F (G&gt;C); L173F (G&gt;T); L173W; L174del; L174M; L174R; F175S; F176L; F177L; F177S; S178P; G183V; E184D; E184G; V191A; I202F; G206A; G206D; G206S; G206V; G209A; G209E; G209R; G209V; S212Y; I213F; I213L; I213T; H214D; H214N; H214Y; G217D; G217R; L219F; L219P; L219R; R220P; Q222H; Q222P; Q222R; Q223R; L226F; L226R; I229F; S230I; S230N; S230R; A231P; A231T; A231V; L232P; M233I (G&gt;A); M233I (G&gt;C); M233L (A&gt;T); M233L (A&gt;C); M233T; M233V; L235P; L235R; L235V; F237I; F237L; I238M; K239N; T245P; A246E; A246P; L248P; L248R; L250F; L2505; L250V; Y2565; A260V; V261F; V261L; L262F; L262V; C263F; C263R; P264L; G266S; P267A; P267L; P267S; R269G; R269H; L271V; V272A; E273A; E273G; T274R; A275V; R278I; R278K; R278S; R278T; E280A; (Paisa); E280G; E280K; L282F; L282R; L282V; F283L; P284L; P284S; A285V; L286P; L286V; T291A; T291P; K311R; E318G; D333G; R352C; R352 S353insR; T354I; R358Q; S365A; S365Y; R377M; R377W; G378E; G378V; G378fs; L381F; L381V; G384A; F386I; F386S; F388L; S390I; S390N; V391F; V391G; L392P; L392V; G394V; A396T; N405S; I408T; A409T; C410Y; V412I; I416T; G417S; L418F; L420R; L424F; L424H; L424R; L424V; A426P; A431E; (Jalisco); A431V; A434C; A434T; L435F; P436Q; P436S; I437V; I439S; I439V; T440del; 869-2A&gt;G; 869-22_869-23ins18 (ΔE9, Δ9, deltaE9); I238_K239insI; S290C; T291_S319del (ΔE9Finn, Δ9Finn, Δ9); S290C; T291_S319del (ΔE9, Δ9); S290C; T291_S319del A&gt;G (ΔE9, Δ9); S290C; T291_S319del G&gt;A (ΔE9, Δ9); S290C; T291_S319del G&gt;T (ΔE9, Δ9); or S290W; S291_R377del (Δ9-10, Delta9-10, p.Ser290_Arg377delinsTrp, g.73671948_73682054del) (mutations are named relative to Uniprot P49768.1/GenBank Ref. No. NM_000021.4), and in PSEN-2; T18M; R29H; G34S; R62C; R62H; P69A; R71W; K82R; A85V; V101M; K115Efs*; T122P; T122R; P123L; E126fs; E126K; S130L; V139M; N141I (Volga German); N141Y; L143H; V1481; K161R; R163H; H169N; M174V; S175C; G212V; V214L; Q228L; Y231C; I235F; A237V; L238F; L238P; M239I; M239V; A252T; A258T; T301M; K306fs; P334A; P334R; P348L; A377V; V393M; T430M; or D439A mutations are named relative to Uniprot P49810.1/GenBank Ref. No. NP_000438.2). See, e.g., Sun et al., Proc Natl Acad Sci USA. 2017; 114:E476-E485; Heilig et al., J Neurosci. 2013 Jul. 10; 33(28):11606-17; Zhou et al., PNAS Nov. 28, 2017 114 (48) 12731-12736. In some embodiments, the methods can include determining that a subject has such a mutation, e.g., using methods known in the art. In some embodiments, the subject has a mutation as described herein (e.g., is identified as having a mutation as described herein using methods known in the art), and optionally has a family history of AD and/or one or more symptoms of AD, and the subject is treated using a method described herein. In some embodiments, the subject does not yet have full AD. 
     Typically, increasing forgetfulness or mild confusion are early symptoms of Alzheimer&#39;s disease. Gradually, cognitive impairment associated with Alzheimer&#39;s disease leads to memory loss, especially recent memories, disorientation and misinterpreting spatial relationships, difficulty in speaking, writing, thinking, reasoning, changes in personality and behavior resulting in depression, anxiety, social withdrawal, mood swings, distrust in others, irritability and aggressiveness, changes in sleeping habits, wandering, loss of inhibitions, delusions, and eventually death. 
     Other Neurodegenerative Diseases, Disorders or Conditions 
     In addition to Alzheimer&#39;s disease, the present methods may be used to treat other neurodegenerative diseases, disorders or conditions, including frontotemporal dementia, various types of memory loss, cognitive impairment including but not limited to mild cognitive impairment (MCI), or other conditions associated with loss of PS1 or PS2, e.g., due to a mutation in PSEN1 or PSEN2, e.g., that creates a dominant negative isoform. 
     Codon-Optimized Presenilin-1 (PSEN1) 
     A codon-optimized presenilin-1 (PSEN1-encoding polynucleotide suitable for use in the compositions and methods described herein can include a full length cDNA or a portion or fragment thereof that encodes a protein retaining substantial gamma secretase activity of the wild-type protein, e.g., at least 50% of the gamma secretase activity, or at least 60, 70, 80, 90, or 95%, or more than 100%, of the activity of the wild-type protein determined by (e.g., in in vitro γ-secretase assays including those described in the Examples section, see also Watanabe et al., J. Neurosci. 32(15):5085-5096 (2012)). In some embodiments, a suitable codon-optimized PSEN1 encodes a protein sequence that is at least 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the full-length wild type PS1 or PS2 protein sequence, respectively. Exemplary wild type genomic, cDNA or protein sequences of human PSEN1/PS1 or PSEN2/PS2 are shown in Table 1 and  FIGS. 4A-C  and  5 A-B. PS1 is normally cleaved into N- and C-terminal fragments that are the active form. PS-1 is processed to give two fragments: an N-terminal 28 kDa fragment, and a C-terminal 18 kDa fragment; the principal endoproteolytic cleavage occurs at and near Met298 in the proximal portion of the large hydrophilic loop (Podlisny et al., Neurobiol Dis. 1997; 3(4):325-37; Marambaud et al., EMBO J. 2002 Apr. 15; 21(8):1948-56). Sequences comprising or encoding these cleaved forms can also be used in the methods and compositions described herein, e.g., encoding amino acids 1-291, 1-292, 1-293, 1-294, 1-295, 1-296, 1-297, 1-298, or 1-299 of SEQ ID NO:5 or a corresponding fragment of SEQ ID NO:6-8. 
     
       
         
           
               
             
               
                 TABLE 1 
               
             
            
               
                   
               
               
                 GenBank Accession Nos. 
               
            
           
           
               
               
               
               
            
               
                 Isoform 
                 mRNA 
                 Protein 
                 RefSeqGene 
               
               
                   
               
               
                 presenilin-1  
                 NM_000021.3 
                 NP_000012.1 
                 NG_007386.2 
               
               
                 isoform I-467 
                 (SEQ ID NO: 1) 
                 (SEQ ID NO: 5) 
                 Range 4965 to 
               
               
                 presenilin-1  
                 NM_007318.2 
                 NP_015557.2 
                 92221 
               
               
                 isoform I-463 
                 (SEQ ID NO: 2) 
                 (SEQ ID NO: 6) 
                   
               
               
                 presenilin-2  
                 NM_000447.2 
                 NP_000438.2 
                 NG_007381.1 
               
               
                 isoform 1 
                 (SEQ ID NO: 3) 
                 (SEQ ID NO: 7) 
                 Range 5001 to 
               
               
                 presenilin-2  
                 NM_012486.2 
                 NP_036618.2 
                 30532 
               
               
                 isoform 2 
                 (SEQ ID NO: 4) 
                 (SEQ ID NO: 8) 
               
               
                   
               
            
           
         
       
     
     
       
         
           
               
               
            
               
                 &gt;NM_000021.3  Homo sapiens  presenilin 1 (PSEN1), transcript variant 
                   
               
               
                 1, mRNA 
               
               
                 (SEQ ID NO: 1) 
                   
               
               
                 AAATGACGACAACGGTGAGGGTTCTCGGGCGGGGCCTGGGACAGGCAGCTCCGGGGTCCGCGGTTTCACA 
                   
               
               
                   
               
               
                 TCGGAAACAAAACAGCGGCTGGTCTGGAAGGAACCTGAGCTACGAGCCGCGGCGGCAGCGGGGCGGCGGG 
               
               
                   
               
               
                 GAAGCGTATACCTAATCTGGGAGCCTGCAAGTGACAACAGCCTTTGCGGTCCTTAGACAGCTTGGCCTGG 
               
               
                   
               
               
                 AGGAGAACACATGAAAGAAAGAACCTCAAGAGGCTTTGTTTTCTGTGAAACAGTATTTCTATACAGTTGC 
               
               
                   
               
               
                 TCCAATGACAGAGTTACCTGCACCGTTGTCCTACTTCCAGAATGCACAGATGTCTGAGGACAACCACCTG 
               
               
                   
               
               
                 AGCAATACTGTACGTAGCCAGAATGACAATAGAGAACGGCAGGAGCACAACGACAGACGGAGCCTTGGCC 
               
               
                   
               
               
                 ACCCTGAGCCATTATCTAATGGACGACCCCAGGGTAACTCCCGGCAGGTGGTGGAGCAAGATGAGGAAGA 
               
               
                   
               
               
                 AGATGAGGAGCTGACATTGAAATATGGCGCCAAGCATGTGATCATGCTCTTTGTCCCTGTGACTCTCTGC 
               
               
                   
               
               
                 ATGGTGGTGGTCGTGGCTACCATTAAGTCAGTCAGCTTTTATACCCGGAAGGATGGGCAGCTAATCTATA 
               
               
                   
               
               
                 CCCCATTCACAGAAGATACCGAGACTGTGGGCCAGAGAGCCCTGCACTCAATTCTGAATGCTGCCATCAT 
               
               
                   
               
               
                 GATCAGTGTCATTGTTGTCATGACTATCCTCCTGGTGGTTCTGTATAAATACAGGTGCTATAAGGTCATC 
               
               
                   
               
               
                 CATGCCTGGCTTATTATATCATCTCTATTGTTGCTGTTCTTTTTTTCATTCATTTACTTGGGGGAAGTGT 
               
               
                   
               
               
                 TTAAAACCTATAACGTTGCTGTGGACTACATTACTGTTGCACTCCTGATCTGGAATTTTGGTGTGGTGGG 
               
               
                   
               
               
                 AATGATTTCCATTCACTGGAAAGGTCCACTTCGACTCCAGCAGGCATATCTCATTATGATTAGTGCCCTC 
               
               
                   
               
               
                 ATGGCCCTGGTGTTTATCAAGTACCTCCCTGAATGGACTGCGTGGCTCATCTTGGCTGTGATTTCAGTAT 
               
               
                   
               
               
                 ATGATTTAGTGGCTGTTTTGTGTCCGAAAGGTCCACTTCGTATGCTGGTTGAAACAGCTCAGGAGAGAAA 
               
               
                   
               
               
                 TGAAACGCTTTTTCCAGCTCTCATTTACTCCTCAACAATGGTGTGGTTGGTGAATATGGCAGAAGGAGAC 
               
               
                   
               
               
                 CCGGAAGCTCAAAGGAGAGTATCCAAAAATTCCAAGTATAATGCAGAAAGCACAGAAAGGGAGTCACAAG 
               
               
                   
               
               
                 ACACTGTTGCAGAGAATGATGATGGCGGGTTCAGTGAGGAATGGGAAGCCCAGAGGGACAGTCATCTAGG 
               
               
                   
               
               
                 GCCTCATCGCTCTACACCTGAGTCACGAGCTGCTGTCCAGGAACTTTCCAGCAGTATCCTCGCTGGTGAA 
               
               
                   
               
               
                 GACCCAGAGGAAAGGGGAGTAAAACTTGGATTGGGAGATTTCATTTTCTACAGTGTTCTGGTTGGTAAAG 
               
               
                   
               
               
                 CCTCAGCAACAGCCAGTGGAGACTGGAACACAACCATAGCCTGTTTCGTAGCCATATTAATTGGTTTGTG 
               
               
                   
               
               
                 CCTTACATTATTACTCCTTGCCATTTTCAAGAAAGCATTGCCAGCTCTTCCAATCTCCATCACCTTTGGG 
               
               
                   
               
               
                 CTTGTTTTCTACTTTGCCACAGATTATCTTGTACAGCCTTTTATGGACCAATTAGCATTCCATCAATTTT 
               
               
                   
               
               
                 ATATCTAGCATATTTGCGGTTAGAATCCCATGGATGTTTCTTCTTTGACTATAACAAAATCTGGGGAGGA 
               
               
                   
               
               
                 CAAAGGTGATTTTCCTGTGTCCACATCTAACAAAGTCAAGATTCCCGGCTGGACTTTTGCAGCTTCCTTC 
               
               
                   
               
               
                 CAAGTCTTCCTGACCACCTTGCACTATTGGACTTTGGAAGGAGGTGCCTATAGAAAACGATTTTGAACAT 
               
               
                   
               
               
                 ACTTCATCGCAGTGGACTGTGTCCCTCGGTGCAGAAACTACCAGATTTGAGGGACGAGGTCAAGGAGATA 
               
               
                   
               
               
                 TGATAGGCCCGGAAGTTGCTGTGCCCCATCAGCAGCTTGACGCGTGGTCACAGGACGATTTCACTGACAC 
               
               
                   
               
               
                 TGCGAACTCTCAGGACTACCGTTACCAAGAGGTTAGGTGAAGTGGTTTAAACCAAACGGAACTCTTCATC 
               
               
                   
               
               
                 TTAAACTACACGTTGAAAATCAACCCAATAATTCTGTATTAACTGAATTCTGAACTTTTCAGGAGGTACT 
               
               
                   
               
               
                 GTGAGGAAGAGCAGGCACCAGCAGCAGAATGGGGAATGGAGAGGTGGGCAGGGGTTCCAGCTTCCCTTTG 
               
               
                   
               
               
                 ATTTTTTGCTGCAGACTCATCCTTTTTAAATGAGACTTGTTTTCCCCTCTCTTTGAGTCAAGTCAAATAT 
               
               
                   
               
               
                 GTAGATTGCCTTTGGCAATTCTTCTTCTCAAGCACTGACACTCATTACCGTCTGTGATTGCCATTTCTTC 
               
               
                   
               
               
                 CCAAGGCCAGTCTGAACCTGAGGTTGCTTTATCCTAAAAGTTTTAACCTCAGGTTCCAAATTCAGTAAAT 
               
               
                   
               
               
                 TTTGGAAACAGTACAGCTATTTCTCATCAATTCTCTATCATGTTGAAGTCAAATTTGGATTTTCCACCAA 
               
               
                   
               
               
                 ATTCTGAATTTGTAGACATACTTGTACGCTCACTTGCCCCAGATGCCTCCTCTGTCCTCATTCTTCTCTC 
               
               
                   
               
               
                 CCACACAAGCAGTCTTTTTCTACAGCCAGTAAGGCAGCTCTGTCGTGGTAGCAGATGGTCCCATTATTCT 
               
               
                   
               
               
                 AGGGTCTTACTCTTTGTATGATGAAAAGAATGTGTTATGAATCGGTGCTGTCAGCCCTGCTGTCAGACCT 
               
               
                   
               
               
                 TCTTCCACAGCAAATGAGATGTATGCCCAAAGACGGTAGAATTAAAGAAGAGTAAAATGGCTGTTGAAGC 
               
               
                   
               
               
                 ACTTTCTGTCCTGGTATTTTGTTTTTGCTTTTGCCACACAGTAGCTCAGAATTTGAACAAATAGCCAAAA 
               
               
                   
               
               
                 GCTGGTGGTTGATGAATTATGAACTAGTTGTATCAACACAAAGCAAGAGTTGGGGAAAGCCATATTTAAC 
               
               
                   
               
               
                 TTGGTGAGCTGTGGGAGAACCTGGTGGCAGAAGGAGAACCAACTGCCAAGGGGAAAGAGAAGGGGCCTCC 
               
               
                   
               
               
                 AGCAGCGAAGGGGATACAGTGAGCTAATGATGTCAAGGAGGAGTTTCAGGTTATTCTCGTCAGCTCCACA 
               
               
                   
               
               
                 AATGGGTGCTTTGTGGTCTCTGCCCGCGTTACCTTTCCTCTCAATGTACCTTTGTGTGAACTGGGCAGTG 
               
               
                   
               
               
                 GAGGTGCCTGCTGCAGTTACCATGGAGTTCAGGCTCTGGGCAGCTCAGTCAGGCAAAACACACAAACAGC 
               
               
                   
               
               
                 CATCAGCCTGTGTGGGCTCAGGGCACCTCTGGACAAAGGCTTGTGGGGCATAACCTTCTTTACCACAGAG 
               
               
                   
               
               
                 AGCCCTTAGCTATGCTGATCAGACCGTAAGCGTTTATGAGAAACTTAGTTTCCTCCTGTGGCTGAGGAGG 
               
               
                   
               
               
                 GGCCAGCTTTTTCTTCTTTTGCCTGCTGTTTTCTCTCCCAATCTATGATATGATATGACCTGGTTTGGGG 
               
               
                   
               
               
                 CTGTCTTTGGTGTTTAGAATATTTGTTTTCTGTCCCAGGATATTTCTTATAAGAACCTAACTTCAAGAGT 
               
               
                   
               
               
                 AGTGTGCGAGTACTGATCTGAATTTAAATTAAAATTGGCTTATATTAGGCAGTCACAGACAGGAAAAATA 
               
               
                   
               
               
                 AGAGCTATGCAAAGAAAGGGGGATTTAAAGTAGTAGGTTCTATCATCTCAATTCATTTTTTTCCATGAAA 
               
               
                   
               
               
                 TCCCTTCTTCCAAGATTCATTCCCTCTCTCAGACATGTGCTAGCATGGGTATTATCATTGAGAAAGCACA 
               
               
                   
               
               
                 GCTACAGCAAAGCCACCTGAATAGCAATTTGTGATTGGAAGCATTCTTGAGGGATCCCTAATCTAGAGTA 
               
               
                   
               
               
                 ATTTATTTGTGTAAGGATCCCAAATGTGTTGCACCTTTCATGATACATTTCTTCTCTGAAGAGGGTACGT 
               
               
                   
               
               
                 GGGGTGTGTGTATTTAAATCCATCCTATGTATTACTGATTGTCCTGTGTAGAAAGATGGCAATTATTCTG 
               
               
                   
               
               
                 TCTCTTTCTCCAAGTTTGAGCCACATCTCAGCCACATTGTTAGACAGTGTACAGAGAACCTATCTTTCCT 
               
               
                   
               
               
                 TTTTTTTTTTTTAAAGGACAGGATTTTGCTGTGTTGCCCAGGCTAGACTTGAACTCCTGGGCTCAAGTAA 
               
               
                   
               
               
                 TCCACCTCAGCCTGAGTAGCTGAGACTACAGCCCATCTTATTTCTTTAAATCATTCATCTCAGGCAGAGA 
               
               
                   
               
               
                 ACTTTTCCCTCAAACATTCTTTTTAGAATTAGTTCAGTCATTCCTAAAACATCCAAATGCTAGTCTTCCA 
               
               
                   
               
               
                 CCATGAAAAATAGATTGTCACTGGAAAGAACAGTAGCAATTTCCATAAGGATGTGCCTTCACTCACACGG 
               
               
                   
               
               
                 GACAGGCGGTGGTTATAGAGTCGGGCAAAACCAGCAGTAGAGTATGACCAGCCAAGCCAATCTGCTTAAT 
               
               
                   
               
               
                 AAAAAGATGGAAGACAGTAAGGAAGGAAAGTAGCCACTAAGAGTCTGAGTCTGACTGGGCTACAGAATAA 
               
               
                   
               
               
                 AGGGTATTTATGGACAGAATGTCATTACATGCCTATGGGAATACCAATCATATTTGGAAGATTTGCAGAT 
               
               
                   
               
               
                 TTTTTTTCAGAGAGGAAAGACTCACCTTCCTGTTTTTGGTTCTCAGTAGGTTCGTGTGTGTTCCTAGAAT 
               
               
                   
               
               
                 CACAGCTCTGACTCCAAATGACTCAATTTCTCAATTAGAAAAAGTAGAAGCTTTCTAAGCAACTTGGAAG 
               
               
                   
               
               
                 AAAACAGTCATAAGTAAGCAATTTGTTGATTTTACTACAGAAGCAACAACTGAAGAGGCAGTGTTTTTAC 
               
               
                   
               
               
                 TTTCAGACTCCGGGATTCCCATTCTGTAGTCTCTCTGCTTTTAAAAACCCTCCTTTTGCAATAGATGCCC 
               
               
                   
               
               
                 AAACAGATGATGTTTATTACTTGTTATTTACGTGGCCTCAGACAGTGTATGTATTCTCGATATAACTTGT 
               
               
                   
               
               
                 AGAGTGTGAAATATAAGTTTAACTACCAAATAAGGTCTCCCAGGGTTAGATGACTGCGGGAAGCCTTTGA 
               
               
                   
               
               
                 TCCCAACCCCCAAGGCTTTGTATATTTGATCATTTGTGATCTAACCCTGGAAGAAAAAGAGCTCAGAAAC 
               
               
                   
               
               
                 CACTATGAAAAAATTTGTTCAGTGTTTTCTGTGTTCCCGTAGGTTCTGGAGTCTGAGGATGCAAAGATGA 
               
               
                   
               
               
                 ATAAGATAAATTCTCAGAATGTAGTTATAATCTCTTGTTTTCTGGTATATGCCATCTTTCTTTAACTTCT 
               
               
                   
               
               
                 CTAAAATATTGGGTATTTGTCAAATAACCACTTTTAACAGTTACCATTACTGAGGGCTTATACATTGGTG 
               
               
                   
               
               
                 TTATAAAAGTGACTTGATTCAGAAATCAATCCATTCAGTAAAGTACTCCTTCTCTAAATTTGCTGTTATG 
               
               
                   
               
               
                 TCTATAAGGAACAGTTTGACCTGCCCTTCTCCTCACCTCCTCACCTGCCTTCCAACATTGAATTTGGAAG 
               
               
                   
               
               
                 GAGACGTGAAAATTGGACATTTGGTTTTGCCCTTGGGCTGGAAACTATCATATAATCATAAGTTTGAGCC 
               
               
                   
               
               
                 TAGAAGTGATCCTTGTGATCTTCTCACCTCTTTAAATTCCCACAACACAAGAGATTAAAAACAGAGGTTT 
               
               
                   
               
               
                 CAGCTCTTCATAGTGCGTTGTGAAATGGCTGGCCAGAGTGTACCAACAAAGCTGTCATCGGGCTCACAGC 
               
               
                   
               
               
                 TCAGAGACATCTGCATGTGATCATCTGCATAGTCCTCTCCTCTAACGGGAAACACCTCAGATTTGCATAT 
               
               
                   
               
               
                 AAAAAAGCACCCTGGTGCTGAAATGAACCCCTTTCTTGAACATCAAAGCTGTCTCCCACAGCCTTGGGCA 
               
               
                   
               
               
                 GCAGGGTGCCTCTTAGTGGATGTGCTGGGTCCACCCTGAGCCCTGACATGTGGTGGCAGCATTGCCAGTT 
               
               
                   
               
               
                 GGTCTGTGTGTCTGTGTAGCAGGGACGATTTCCCAGAAAGCAATTTTCCTTTTGAAATACGTAATTGTTG 
               
               
                   
               
               
                 AGACTAGGCAGTTTCAAAGTCAGCTGCATATAGTAGCAAGTACAGGACTGTCTTGTTTTTGGTGTCCTTG 
               
               
                   
               
               
                 GAGGTGCTGGGGTGAGGGTTTCAGTGGGATCATTTACTCTCACATGTTGTCTGCCTTCTGCTTCTGTGGA 
               
               
                   
               
               
                 CACTGCTTTGTACTTAATTCAGACAGACTGTGAATACACCTTTTTTATAAATACCTTTCAAATTCTTGGT 
               
               
                   
               
               
                 AAGATATAATTTTGATAGCTGATTGCAGATTTTCTGTATTTGTCAGATTAATAAAGACTGCATGAATCCA 
               
               
                   
               
               
                 AAAAAAAAAAAAAAAAA 
               
               
                   
               
               
                 &gt;NM_007318.2  Homo sapiens  presenilin 1 (PSEN1), transcript variant 
               
               
                 2, mRNA 
               
               
                 (SEQ ID NO: 2) 
                   
               
               
                 AAATGACGACAACGGTGAGGGTTCTCGGGCGGGGCCTGGGACAGGCAGCTCCGGGGTCCGCGGTTTCACA 
                   
               
               
                   
               
               
                 TCGGAAACAAAACAGCGGCTGGTCTGGAAGGAACCTGAGCTACGAGCCGCGGCGGCAGCGGGGCGGCGGG 
               
               
                   
               
               
                 GAAGCGTATACCTAATCTGGGAGCCTGCAAGTGACAACAGCCTTTGCGGTCCTTAGACAGCTTGGCCTGG 
               
               
                   
               
               
                 AGGAGAACACATGAAAGAAAGAACCTCAAGAGGCTTTGTTTTCTGTGAAACAGTATTTCTATACAGTTGC 
               
               
                   
               
               
                 TCCAATGACAGAGTTACCTGCACCGTTGTCCTACTTCCAGAATGCACAGATGTCTGAGGACAACCACCTG 
               
               
                   
               
               
                 AGCAATACTAATGACAATAGAGAACGGCAGGAGCACAACGACAGACGGAGCCTTGGCCACCCTGAGCCAT 
               
               
                   
               
               
                 TATCTAATGGACGACCCCAGGGTAACTCCCGGCAGGTGGTGGAGCAAGATGAGGAAGAAGATGAGGAGCT 
               
               
                   
               
               
                 GACATTGAAATATGGCGCCAAGCATGTGATCATGCTCTTTGTCCCTGTGACTCTCTGCATGGTGGTGGTC 
               
               
                   
               
               
                 GTGGCTACCATTAAGTCAGTCAGCTTTTATACCCGGAAGGATGGGCAGCTAATCTATACCCCATTCACAG 
               
               
                   
               
               
                 AAGATACCGAGACTGTGGGCCAGAGAGCCCTGCACTCAATTCTGAATGCTGCCATCATGATCAGTGTCAT 
               
               
                   
               
               
                 TGTTGTCATGACTATCCTCCTGGTGGTTCTGTATAAATACAGGTGCTATAAGGTCATCCATGCCTGGCTT 
               
               
                   
               
               
                 ATTATATCATCTCTATTGTTGCTGTTCTTTTTTTCATTCATTTACTTGGGGGAAGTGTTTAAAACCTATA 
               
               
                   
               
               
                 ACGTTGCTGTGGACTACATTACTGTTGCACTCCTGATCTGGAATTTTGGTGTGGTGGGAATGATTTCCAT 
               
               
                   
               
               
                 TCACTGGAAAGGTCCACTTCGACTCCAGCAGGCATATCTCATTATGATTAGTGCCCTCATGGCCCTGGTG 
               
               
                   
               
               
                 TTTATCAAGTACCTCCCTGAATGGACTGCGTGGCTCATCTTGGCTGTGATTTCAGTATATGATTTAGTGG 
               
               
                   
               
               
                 CTGTTTTGTGTCCGAAAGGTCCACTTCGTATGCTGGTTGAAACAGCTCAGGAGAGAAATGAAACGCTTTT 
               
               
                   
               
               
                 TCCAGCTCTCATTTACTCCTCAACAATGGTGTGGTTGGTGAATATGGCAGAAGGAGACCCGGAAGCTCAA 
               
               
                   
               
               
                 AGGAGAGTATCCAAAAATTCCAAGTATAATGCAGAAAGCACAGAAAGGGAGTCACAAGACACTGTTGCAG 
               
               
                   
               
               
                 AGAATGATGATGGCGGGTTCAGTGAGGAATGGGAAGCCCAGAGGGACAGTCATCTAGGGCCTCATCGCTC 
               
               
                   
               
               
                 TACACCTGAGTCACGAGCTGCTGTCCAGGAACTTTCCAGCAGTATCCTCGCTGGTGAAGACCCAGAGGAA 
               
               
                   
               
               
                 AGGGGAGTAAAACTTGGATTGGGAGATTTCATTTTCTACAGTGTTCTGGTTGGTAAAGCCTCAGCAACAG 
               
               
                   
               
               
                 CCAGTGGAGACTGGAACACAACCATAGCCTGTTTCGTAGCCATATTAATTGGTTTGTGCCTTACATTATT 
               
               
                   
               
               
                 ACTCCTTGCCATTTTCAAGAAAGCATTGCCAGCTCTTCCAATCTCCATCACCTTTGGGCTTGTTTTCTAC 
               
               
                   
               
               
                 TTTGCCACAGATTATCTTGTACAGCCTTTTATGGACCAATTAGCATTCCATCAATTTTATATCTAGCATA 
               
               
                   
               
               
                 TTTGCGGTTAGAATCCCATGGATGTTTCTTCTTTGACTATAACAAAATCTGGGGAGGACAAAGGTGATTT 
               
               
                   
               
               
                 TCCTGTGTCCACATCTAACAAAGTCAAGATTCCCGGCTGGACTTTTGCAGCTTCCTTCCAAGTCTTCCTG 
               
               
                   
               
               
                 ACCACCTTGCACTATTGGACTTTGGAAGGAGGTGCCTATAGAAAACGATTTTGAACATACTTCATCGCAG 
               
               
                   
               
               
                 TGGACTGTGTCCCTCGGTGCAGAAACTACCAGATTTGAGGGACGAGGTCAAGGAGATATGATAGGCCCGG 
               
               
                   
               
               
                 AAGTTGCTGTGCCCCATCAGCAGCTTGACGCGTGGTCACAGGACGATTTCACTGACACTGCGAACTCTCA 
               
               
                   
               
               
                 GGACTACCGTTACCAAGAGGTTAGGTGAAGTGGTTTAAACCAAACGGAACTCTTCATCTTAAACTACACG 
               
               
                   
               
               
                 TTGAAAATCAACCCAATAATTCTGTATTAACTGAATTCTGAACTTTTCAGGAGGTACTGTGAGGAAGAGC 
               
               
                   
               
               
                 AGGCACCAGCAGCAGAATGGGGAATGGAGAGGTGGGCAGGGGTTCCAGCTTCCCTTTGATTTTTTGCTGC 
               
               
                   
               
               
                 AGACTCATCCTTTTTAAATGAGACTTGTTTTCCCCTCTCTTTGAGTCAAGTCAAATATGTAGATTGCCTT 
               
               
                   
               
               
                 TGGCAATTCTTCTTCTCAAGCACTGACACTCATTACCGTCTGTGATTGCCATTTCTTCCCAAGGCCAGTC 
               
               
                   
               
               
                 TGAACCTGAGGTTGCTTTATCCTAAAAGTTTTAACCTCAGGTTCCAAATTCAGTAAATTTTGGAAACAGT 
               
               
                   
               
               
                 ACAGCTATTTCTCATCAATTCTCTATCATGTTGAAGTCAAATTTGGATTTTCCACCAAATTCTGAATTTG 
               
               
                   
               
               
                 TAGACATACTTGTACGCTCACTTGCCCCAGATGCCTCCTCTGTCCTCATTCTTCTCTCCCACACAAGCAG 
               
               
                   
               
               
                 TCTTTTTCTACAGCCAGTAAGGCAGCTCTGTCGTGGTAGCAGATGGTCCCATTATTCTAGGGTCTTACTC 
               
               
                   
               
               
                 TTTGTATGATGAAAAGAATGTGTTATGAATCGGTGCTGTCAGCCCTGCTGTCAGACCTTCTTCCACAGCA 
               
               
                   
               
               
                 AATGAGATGTATGCCCAAAGACGGTAGAATTAAAGAAGAGTAAAATGGCTGTTGAAGCACTTTCTGTCCT 
               
               
                   
               
               
                 GGTATTTTGTTTTTGCTTTTGCCACACAGTAGCTCAGAATTTGAACAAATAGCCAAAAGCTGGTGGTTGA 
               
               
                   
               
               
                 TGAATTATGAACTAGTTGTATCAACACAAAGCAAGAGTTGGGGAAAGCCATATTTAACTTGGTGAGCTGT 
               
               
                   
               
               
                 GGGAGAACCTGGTGGCAGAAGGAGAACCAACTGCCAAGGGGAAAGAGAAGGGGCCTCCAGCAGCGAAGGG 
               
               
                   
               
               
                 GATACAGTGAGCTAATGATGTCAAGGAGGAGTTTCAGGTTATTCTCGTCAGCTCCACAAATGGGTGCTTT 
               
               
                   
               
               
                 GTGGTCTCTGCCCGCGTTACCTTTCCTCTCAATGTACCTTTGTGTGAACTGGGCAGTGGAGGTGCCTGCT 
               
               
                   
               
               
                 GCAGTTACCATGGAGTTCAGGCTCTGGGCAGCTCAGTCAGGCAAAACACACAAACAGCCATCAGCCTGTG 
               
               
                   
               
               
                 TGGGCTCAGGGCACCTCTGGACAAAGGCTTGTGGGGCATAACCTTCTTTACCACAGAGAGCCCTTAGCTA 
               
               
                   
               
               
                 TGCTGATCAGACCGTAAGCGTTTATGAGAAACTTAGTTTCCTCCTGTGGCTGAGGAGGGGCCAGCTTTTT 
               
               
                   
               
               
                 CTTCTTTTGCCTGCTGTTTTCTCTCCCAATCTATGATATGATATGACCTGGTTTGGGGCTGTCTTTGGTG 
               
               
                   
               
               
                 TTTAGAATATTTGTTTTCTGTCCCAGGATATTTCTTATAAGAACCTAACTTCAAGAGTAGTGTGCGAGTA 
               
               
                   
               
               
                 CTGATCTGAATTTAAATTAAAATTGGCTTATATTAGGCAGTCACAGACAGGAAAAATAAGAGCTATGCAA 
               
               
                   
               
               
                 AGAAAGGGGGATTTAAAGTAGTAGGTTCTATCATCTCAATTCATTTTTTTCCATGAAATCCCTTCTTCCA 
               
               
                   
               
               
                 AGATTCATTCCCTCTCTCAGACATGTGCTAGCATGGGTATTATCATTGAGAAAGCACAGCTACAGCAAAG 
               
               
                   
               
               
                 CCACCTGAATAGCAATTTGTGATTGGAAGCATTCTTGAGGGATCCCTAATCTAGAGTAATTTATTTGTGT 
               
               
                   
               
               
                 AAGGATCCCAAATGTGTTGCACCTTTCATGATACATTTCTTCTCTGAAGAGGGTACGTGGGGTGTGTGTA 
               
               
                   
               
               
                 TTTAAATCCATCCTATGTATTACTGATTGTCCTGTGTAGAAAGATGGCAATTATTCTGTCTCTTTCTCCA 
               
               
                   
               
               
                 AGTTTGAGCCACATCTCAGCCACATTGTTAGACAGTGTACAGAGAACCTATCTTTCCTTTTTTTTTTTTT 
               
               
                   
               
               
                 AAAGGACAGGATTTTGCTGTGTTGCCCAGGCTAGACTTGAACTCCTGGGCTCAAGTAATCCACCTCAGCC 
               
               
                   
               
               
                 TGAGTAGCTGAGACTACAGCCCATCTTATTTCTTTAAATCATTCATCTCAGGCAGAGAACTTTTCCCTCA 
               
               
                   
               
               
                 AACATTCTTTTTAGAATTAGTTCAGTCATTCCTAAAACATCCAAATGCTAGTCTTCCACCATGAAAAATA 
               
               
                   
               
               
                 GATTGTCACTGGAAAGAACAGTAGCAATTTCCATAAGGATGTGCCTTCACTCACACGGGACAGGCGGTGG 
               
               
                   
               
               
                 TTATAGAGTCGGGCAAAACCAGCAGTAGAGTATGACCAGCCAAGCCAATCTGCTTAATAAAAAGATGGAA 
               
               
                   
               
               
                 GACAGTAAGGAAGGAAAGTAGCCACTAAGAGTCTGAGTCTGACTGGGCTACAGAATAAAGGGTATTTATG 
               
               
                   
               
               
                 GACAGAATGTCATTACATGCCTATGGGAATACCAATCATATTTGGAAGATTTGCAGATTTTTTTTCAGAG 
               
               
                   
               
               
                 AGGAAAGACTCACCTTCCTGTTTTTGGTTCTCAGTAGGTTCGTGTGTGTTCCTAGAATCACAGCTCTGAC 
               
               
                   
               
               
                 TCCAAATGACTCAATTTCTCAATTAGAAAAAGTAGAAGCTTTCTAAGCAACTTGGAAGAAAACAGTCATA 
               
               
                   
               
               
                 AGTAAGCAATTTGTTGATTTTACTACAGAAGCAACAACTGAAGAGGCAGTGTTTTTACTTTCAGACTCCG 
               
               
                   
               
               
                 GGATTCCCATTCTGTAGTCTCTCTGCTTTTAAAAACCCTCCTTTTGCAATAGATGCCCAAACAGATGATG 
               
               
                   
               
               
                 TTTATTACTTGTTATTTACGTGGCCTCAGACAGTGTATGTATTCTCGATATAACTTGTAGAGTGTGAAAT 
               
               
                   
               
               
                 ATAAGTTTAACTACCAAATAAGGTCTCCCAGGGTTAGATGACTGCGGGAAGCCTTTGATCCCAACCCCCA 
               
               
                   
               
               
                 AGGCTTTGTATATTTGATCATTTGTGATCTAACCCTGGAAGAAAAAGAGCTCAGAAACCACTATGAAAAA 
               
               
                   
               
               
                 ATTTGTTCAGTGTTTTCTGTGTTCCCGTAGGTTCTGGAGTCTGAGGATGCAAAGATGAATAAGATAAATT 
               
               
                   
               
               
                 CTCAGAATGTAGTTATAATCTCTTGTTTTCTGGTATATGCCATCTTTCTTTAACTTCTCTAAAATATTGG 
               
               
                   
               
               
                 GTATTTGTCAAATAACCACTTTTAACAGTTACCATTACTGAGGGCTTATACATTGGTGTTATAAAAGTGA 
               
               
                   
               
               
                 CTTGATTCAGAAATCAATCCATTCAGTAAAGTACTCCTTCTCTAAATTTGCTGTTATGTCTATAAGGAAC 
               
               
                   
               
               
                 AGTTTGACCTGCCCTTCTCCTCACCTCCTCACCTGCCTTCCAACATTGAATTTGGAAGGAGACGTGAAAA 
               
               
                   
               
               
                 TTGGACATTTGGTTTTGCCCTTGGGCTGGAAACTATCATATAATCATAAGTTTGAGCCTAGAAGTGATCC 
               
               
                   
               
               
                 TTGTGATCTTCTCACCTCTTTAAATTCCCACAACACAAGAGATTAAAAACAGAGGTTTCAGCTCTTCATA 
               
               
                   
               
               
                 GTGCGTTGTGAAATGGCTGGCCAGAGTGTACCAACAAAGCTGTCATCGGGCTCACAGCTCAGAGACATCT 
               
               
                   
               
               
                 GCATGTGATCATCTGCATAGTCCTCTCCTCTAACGGGAAACACCTCAGATTTGCATATAAAAAAGCACCC 
               
               
                   
               
               
                 TGGTGCTGAAATGAACCCCTTTCTTGAACATCAAAGCTGTCTCCCACAGCCTTGGGCAGCAGGGTGCCTC 
               
               
                   
               
               
                 TTAGTGGATGTGCTGGGTCCACCCTGAGCCCTGACATGTGGTGGCAGCATTGCCAGTTGGTCTGTGTGTC 
               
               
                   
               
               
                 TGTGTAGCAGGGACGATTTCCCAGAAAGCAATTTTCCTTTTGAAATACGTAATTGTTGAGACTAGGCAGT 
               
               
                   
               
               
                 TTCAAAGTCAGCTGCATATAGTAGCAAGTACAGGACTGTCTTGTTTTTGGTGTCCTTGGAGGTGCTGGGG 
               
               
                   
               
               
                 TGAGGGTTTCAGTGGGATCATTTACTCTCACATGTTGTCTGCCTTCTGCTTCTGTGGACACTGCTTTGTA 
               
               
                   
               
               
                 CTTAATTCAGACAGACTGTGAATACACCTTTTTTATAAATACCTTTCAAATTCTTGGTAAGATATAATTT 
               
               
                   
               
               
                 TGATAGCTGATTGCAGATTTTCTGTATTTGTCAGATTAATAAAGACTGCATGAATCCAAAAAAAAAAAAA 
               
               
                   
               
               
                 AAAAA 
               
               
                   
               
               
                 &gt;NM_000447.2  Homo sapiens  presenilin 2 (PSEN2), transcript variant 
               
               
                 1, mRNA 
               
               
                 (SEQ ID NO: 3) 
                   
               
               
                 GGGGCCTGGGCCGGCGCCGGGTCCGGCCGGGCGCTCAGCCAGCTGCGTAAACTCCGCTGGAGCGCGGCGG 
                   
               
               
                   
               
               
                 CAGAGCAGGCATTTCCAGCAGTGAGGAGACAGCCAGAAGCAAGCTTTTGGAGCTGAAGGAACCTGAGACA 
               
               
                   
               
               
                 GAAGCTAGTCCCCCCTCTGAATTTTACTGATGAAGAAACTGAGGCCACAGAGCTAAAGTGACTTTTCCCA 
               
               
                   
               
               
                 AGGTCGCCCAGCGAGGACGTGGGACTTCTCAGACGTCAGGAGAGTGATGTGAGGGAGCTGTGTGACCATA 
               
               
                   
               
               
                 GAAAGTGACGTGTTAAAAACCAGCGCTGCCCTCTTTGAAAGCCAGGGAGCATCATTCATTTAGCCTGCTG 
               
               
                   
               
               
                 AGAAGAAGAAACCAAGTGTCCGGGATTCAGACCTCTCTGCGGCCCCAAGTGTTCGTGGTGCTTCCAGAGG 
               
               
                   
               
               
                 CAGGGCTATGCTCACATTCATGGCCTCTGACAGCGAGGAAGAAGTGTGTGATGAGCGGACGTCCCTAATG 
               
               
                   
               
               
                 TCGGCTGAGAGCCCCACGCCGCGCTCCTGCCAGGAGGGCAGGCAGGGCCCAGAGGATGGAGAGAACACTG 
               
               
                   
               
               
                 CCCAGTGGAGAAGCCAGGAGAACGAGGAGGACGGTGAGGAGGACCCTGACCGCTATGTCTGTAGTGGGGT 
               
               
                   
               
               
                 TCCCGGGCGGCCGCCAGGCCTGGAGGAAGAGCTGACCCTCAAATACGGAGCGAAGCACGTGATCATGCTG 
               
               
                   
               
               
                 TTTGTGCCTGTCACTCTGTGCATGATCGTGGTGGTAGCCACCATCAAGTCTGTGCGCTTCTACACAGAGA 
               
               
                   
               
               
                 AGAATGGACAGCTCATCTACACGCCATTCACTGAGGACACACCCTCGGTGGGCCAGCGCCTCCTCAACTC 
               
               
                   
               
               
                 CGTGCTGAACACCCTCATCATGATCAGCGTCATCGTGGTTATGACCATCTTCTTGGTGGTGCTCTACAAG 
               
               
                   
               
               
                 TACCGCTGCTACAAGTTCATCCATGGCTGGTTGATCATGTCTTCACTGATGCTGCTGTTCCTCTTCACCT 
               
               
                   
               
               
                 ATATCTACCTTGGGGAAGTGCTCAAGACCTACAATGTGGCCATGGACTACCCCACCCTCTTGCTGACTGT 
               
               
                   
               
               
                 CTGGAACTTCGGGGCAGTGGGCATGGTGTGCATCCACTGGAAGGGCCCTCTGGTGCTGCAGCAGGCCTAC 
               
               
                   
               
               
                 CTCATCATGATCAGTGCGCTCATGGCCCTAGTGTTCATCAAGTACCTCCCAGAGTGGTCCGCGTGGGTCA 
               
               
                   
               
               
                 TCCTGGGCGCCATCTCTGTGTATGATCTCGTGGCTGTGCTGTGTCCCAAAGGGCCTCTGAGAATGCTGGT 
               
               
                   
               
               
                 AGAAACTGCCCAGGAGAGAAATGAGCCCATATTCCCTGCCCTGATATACTCATCTGCCATGGTGTGGACG 
               
               
                   
               
               
                 GTTGGCATGGCGAAGCTGGACCCCTCCTCTCAGGGTGCCCTCCAGCTCCCCTACGACCCGGAGATGGAAG 
               
               
                   
               
               
                 AAGACTCCTATGACAGTTTTGGGGAGCCTTCATACCCCGAAGTCTTTGAGCCTCCCTTGACTGGCTACCC 
               
               
                   
               
               
                 AGGGGAGGAGCTGGAGGAAGAGGAGGAAAGGGGCGTGAAGCTTGGCCTCGGGGACTTCATCTTCTACAGT 
               
               
                   
               
               
                 GTGCTGGTGGGCAAGGCGGCTGCCACGGGCAGCGGGGACTGGAATACCACGCTGGCCTGCTTCGTGGCCA 
               
               
                   
               
               
                 TCCTCATTGGCTTGTGTCTGACCCTCCTGCTGCTTGCTGTGTTCAAGAAGGCGCTGCCCGCCCTCCCCAT 
               
               
                   
               
               
                 CTCCATCACGTTCGGGCTCATCTTTTACTTCTCCACGGACAACCTGGTGCGGCCGTTCATGGACACCCTG 
               
               
                   
               
               
                 GCCTCCCATCAGCTCTACATCTGAGGGACATGGTGTGCCACAGGCTGCAAGCTGCAGGGAATTTTCATTG 
               
               
                   
               
               
                 GATGCAGTTGTATAGTTTTACACTCTAGTGCCATATATTTTTAAGACTTTTCTTTCCTTAAAAAATAAAG 
               
               
                   
               
               
                 TACGTGTTTACTTGGTGAGGAGGAGGCAGAACCAGCTCTTTGGTGCCAGCTGTTTCATCACCAGACTTTG 
               
               
                   
               
               
                 GCTCCCGCTTTGGGGAGCGCCTCGCTTCACGGACAGGAAGCACAGCAGGTTTATCCAGATGAACTGAGAA 
               
               
                   
               
               
                 GGTCAGATTAGGGCGGGGAGAAGAGCATCCGGCATGAGGGCTGAGATGCGCAAAGAGTGTGCTCGGGAGT 
               
               
                   
               
               
                 GGCCCCTGGCACCTGGGTGCTCTGGCTGGAGAGGAAAAGCCAGTTCCCTACGAGGAGTGTTCCCAATGCT 
               
               
                   
               
               
                 TTGTCCATGATGTCCTTGTTATTTTATTGCCTTTAGAAACTGAGTCCTGTTCTTGTTACGGCAGTCACAC 
               
               
                   
               
               
                 TGCTGGGAAGTGGCTTAATAGTAATATCAATAAATAGATGAGTCCTGTTAGAATCTTGAAAA 
               
               
                   
               
               
                 &gt;NM_012486.2  Homo sapiens  presenilin 2 (PSEN2), transcript variant 
               
               
                 2, mRNA 
               
               
                 (SEQ ID NO: 4) 
                   
               
               
                 GGGGCCTGGGCCGGCGCCGGGTCCGGCCGGGCGCTCAGCCAGCTGCGTAAACTCCGCTGGAGCGCGGCGG 
                   
               
               
                   
               
               
                 CAGAGCAGGCATTTCCAGCAGTGAGGAGACAGCCAGAAGCAAGCTTTTGGAGCTGAAGGAACCTGAGACA 
               
               
                   
               
               
                 GAAGCTAGTCCCCCCTCTGAATTTTACTGATGAAGAAACTGAGGCCACAGAGCTAAAGTGACTTTTCCCA 
               
               
                   
               
               
                 AGGTCGCCCAGCGAGGACGTGGGACTTCTCAGACGTCAGGAGAGTGATGTGAGGGAGCTGTGTGACCATA 
               
               
                   
               
               
                 GAAAGTGACGTGTTAAAAACCAGCGCTGCCCTCTTTGAAAGCCAGGGAGCATCATTCATTTAGCCTGCTG 
               
               
                   
               
               
                 AGAAGAAGAAACCAAGTGTCCGGGATTCAGACCTCTCTGCGGCCCCAAGTGTTCGTGGTGCTTCCAGAGG 
               
               
                   
               
               
                 CAGGGCTATGCTCACATTCATGGCCTCTGACAGCGAGGAAGAAGTGTGTGATGAGCGGACGTCCCTAATG 
               
               
                   
               
               
                 TCGGCTGAGAGCCCCACGCCGCGCTCCTGCCAGGAGGGCAGGCAGGGCCCAGAGGATGGAGAGAACACTG 
               
               
                   
               
               
                 CCCAGTGGAGAAGCCAGGAGAACGAGGAGGACGGTGAGGAGGACCCTGACCGCTATGTCTGTAGTGGGGT 
               
               
                   
               
               
                 TCCCGGGCGGCCGCCAGGCCTGGAGGAAGAGCTGACCCTCAAATACGGAGCGAAGCACGTGATCATGCTG 
               
               
                   
               
               
                 TTTGTGCCTGTCACTCTGTGCATGATCGTGGTGGTAGCCACCATCAAGTCTGTGCGCTTCTACACAGAGA 
               
               
                   
               
               
                 AGAATGGACAGCTCATCTACACGCCATTCACTGAGGACACACCCTCGGTGGGCCAGCGCCTCCTCAACTC 
               
               
                   
               
               
                 CGTGCTGAACACCCTCATCATGATCAGCGTCATCGTGGTTATGACCATCTTCTTGGTGGTGCTCTACAAG 
               
               
                   
               
               
                 TACCGCTGCTACAAGTTCATCCATGGCTGGTTGATCATGTCTTCACTGATGCTGCTGTTCCTCTTCACCT 
               
               
                   
               
               
                 ATATCTACCTTGGGGAAGTGCTCAAGACCTACAATGTGGCCATGGACTACCCCACCCTCTTGCTGACTGT 
               
               
                   
               
               
                 CTGGAACTTCGGGGCAGTGGGCATGGTGTGCATCCACTGGAAGGGCCCTCTGGTGCTGCAGCAGGCCTAC 
               
               
                   
               
               
                 CTCATCATGATCAGTGCGCTCATGGCCCTAGTGTTCATCAAGTACCTCCCAGAGTGGTCCGCGTGGGTCA 
               
               
                   
               
               
                 TCCTGGGCGCCATCTCTGTGTATGATCTCGTGGCTGTGCTGTGTCCCAAAGGGCCTCTGAGAATGCTGGT 
               
               
                   
               
               
                 AGAAACTGCCCAGGAGAGAAATGAGCCCATATTCCCTGCCCTGATATACTCATCTGCCATGGTGTGGACG 
               
               
                   
               
               
                 GTTGGCATGGCGAAGCTGGACCCCTCCTCTCAGGGTGCCCTCCAGCTCCCCTACGACCCGGAGATGGAAG 
               
               
                   
               
               
                 ACTCCTATGACAGTTTTGGGGAGCCTTCATACCCCGAAGTCTTTGAGCCTCCCTTGACTGGCTACCCAGG 
               
               
                   
               
               
                 GGAGGAGCTGGAGGAAGAGGAGGAAAGGGGCGTGAAGCTTGGCCTCGGGGACTTCATCTTCTACAGTGTG 
               
               
                   
               
               
                 CTGGTGGGCAAGGCGGCTGCCACGGGCAGCGGGGACTGGAATACCACGCTGGCCTGCTTCGTGGCCATCC 
               
               
                   
               
               
                 TCATTGGCTTGTGTCTGACCCTCCTGCTGCTTGCTGTGTTCAAGAAGGCGCTGCCCGCCCTCCCCATCTC 
               
               
                   
               
               
                 CATCACGTTCGGGCTCATCTTTTACTTCTCCACGGACAACCTGGTGCGGCCGTTCATGGACACCCTGGCC 
               
               
                   
               
               
                 TCCCATCAGCTCTACATCTGAGGGACATGGTGTGCCACAGGCTGCAAGCTGCAGGGAATTTTCATTGGAT 
               
               
                   
               
               
                 GCAGTTGTATAGTTTTACACTCTAGTGCCATATATTTTTAAGACTTTTCTTTCCTTAAAAAATAAAGTAC 
               
               
                   
               
               
                 GTGTTTACTTGGTGAGGAGGAGGCAGAACCAGCTCTTTGGTGCCAGCTGTTTCATCACCAGACTTTGGCT 
               
               
                   
               
               
                 CCCGCTTTGGGGAGCGCCTCGCTTCACGGACAGGAAGCACAGCAGGTTTATCCAGATGAACTGAGAAGGT 
               
               
                   
               
               
                 CAGATTAGGGCGGGGAGAAGAGCATCCGGCATGAGGGCTGAGATGCGCAAAGAGTGTGCTCGGGAGTGGC 
               
               
                   
               
               
                 CCCTGGCACCTGGGTGCTCTGGCTGGAGAGGAAAAGCCAGTTCCCTACGAGGAGTGTTCCCAATGCTTTG 
               
               
                   
               
               
                 TCCATGATGTCCTTGTTATTTTATTGCCTTTAGAAACTGAGTCCTGTTCTTGTTACGGCAGTCACACTGC 
               
               
                   
               
               
                 TGGGAAGTGGCTTAATAGTAATATCAATAAATAGATGAGTCCTGTTAGAATCTTGAAAA 
               
               
                   
               
               
                 &gt;NP_000012.1 presenilin-1 isoform 1-467 [ Homo sapiens ] 
               
               
                 (SEQ ID NO: 5) 
                   
               
               
                 MTELPAPLSYFQNAQMSEDNHLSNTVRSQNDNRERQEHNDRRSLGHPEPLSNGRPQGNSRQVVEQ 
                   
               
               
                   
               
               
                 DEEEDEELTLKYGAKHVIMLFVPVTLCMVVVVATIKSVSFYTRKDGQLIYTPFTEDTETVGQRAL 
               
               
                   
               
               
                 HSILNAAIMISVIVVMTILLVVLYKYRCYKVIHAWLIISSLLLLFFFSFIYLGEVFKTYN 
               
               
                   
               
               
                 VAVDYITVALLIWNFGVVGMISIHWKGPLRLQQAYLIMISALMALVFIKYLPEWTAWLI 
               
               
                   
               
               
                 LAVISVYDLVAVLCPKGPLRMLVETAQERNETLFPALIYSSTMVWLVNMAEGDPEAQRR 
               
               
                   
               
               
                 VSKNSKYNAESTERESQDTVAENDDGGFSEEWEAQRDSHLGPHRSTPESRAAVQELSSS 
               
               
                   
               
               
                 ILAGEDPEERGVKLGLGDFIFYSVLVGKASATASGDWNTTIACFVAILIGLCLTLLLLA 
               
               
                   
               
               
                 IFKKALPALPISITFGLVFYFATDYLVQPFMDQLAFHQFYI 
               
               
                   
               
               
                 &gt;NP_015557.2 presenilin-1 isoform 1-463 [Homo sapiens] 
               
               
                 (SEQ ID NO: 6) 
                   
               
               
                 MTELPAPLSYFQNAQMSEDNHLSNTNDNRERQEHNDRRSLGHPEPLSNGRPQGNSRQVVEQDEEEDEE 
                   
               
               
                   
               
               
                 LTLKYGAKHVIMLFVPVTLCMVVVVATIKSVSFYTRKDGQLIYTPFTEDTETVGQRALHSILNAAIMI 
               
               
                   
               
               
                 SVIVVMTILLVVLYKYRCYKVIHAWLIISSLLLLFFFSFIYLGEVFKTYNVAVDYITVALLIWNFGVV 
               
               
                   
               
               
                 GMISIHWKGPLRLQQAYLIMISALMALVFIKYLPEWTAWLILAVISVYDLVAVLCPKGPLRMLVETAQ 
               
               
                   
               
               
                 ERNETLFPALIYSSTMVWLVNMAEGDPEAQRRVSKNSKYNAESTERESQDTVAENDDGGFSEEWEAQR 
               
               
                   
               
               
                 DSHLGPHRSTPESRAAVQELSSSILAGEDPEERGVKLGLGDFIFYSVLVGKASATASGDWNTTIACFV 
               
               
                   
               
               
                 AILIGLCLTLLLLAIFKKALPALPISITFGLVFYFATDYLVQPFMDQLAFHQFYI 
               
               
                   
               
               
                 &gt;NP_000438.2 presenilin-2 isoform 1 [ Homo sapiens ] 
               
               
                 (SEQ ID NO: 7) 
                   
               
               
                 MLTFMASDSEEEVCDERTSLMSAESPTPRSCQEGRQGPEDGENTAQWRSQENEEDGEEDPDRYVCSGV 
                   
               
               
                   
               
               
                 PGRPPGLEEELTLKYGAKHVIMLFVPVTLCMIVVVATIKSVRFYTEKNGQLIYTPFTEDTPSVGQRLL 
               
               
                   
               
               
                 NSVLNTLIMISVIVVMTIFLVVLYKYRCYKFIHGWLIMSSLMLLFLFTYIYLGEVLKTYNVAMDYPTL 
               
               
                   
               
               
                 LLTVWNFGAVGMVCIHWKGPLVLQQAYLIMISALMALVFIKYLPEWSAWVILGAISVYDLVAVLCPKG 
               
               
                   
               
               
                 PLRMLVETAQERNEPIFPALIYSSAMVWTVGMAKLDPSSQGALQLPYDPEMEEDSYDSFGEPSYPEVF 
               
               
                   
               
               
                 EPPLTGYPGEELEEEEERGVKLGLGDFIFYSVLVGKAAATGSGDWNTTLACFVAILIGLCLTLLLLAV 
               
               
                   
               
               
                 FKKALPALPISITEGLIFYFSTDNLVRPFMDTLASHQLYI 
               
               
                   
               
               
                 &gt;NP_036618.2 presenilin-2 isoform 2 [ Homo sapiens ] 
               
               
                 (SEQ ID NO: 8) 
                   
               
               
                 MLTFMASDSEEEVCDERTSLMSAESPTPRSCQEGRQGPEDGENTAQWRSQENEEDGEEDPDRYVCSGV 
                   
               
               
                   
               
               
                 PGRPPGLEEELTLKYGAKHVIMLFVPVTLCMIVVVATIKSVRFYTEKNGQLIYTPFTEDTPSVGQRLL 
               
               
                   
               
               
                 NSVLNTLIMISVIVVMTIFLVVLYKYRCYKFIHGWLIMSSLMLLFLFTYIYLGEVLKTYNVAMDYPTL 
               
               
                   
               
               
                 LLTVWNFGAVGMVCIHWKGPLVLQQAYLIMISALMALVFIKYLPEWSAWVILGAISVYDLVAVLCPKG 
               
               
                   
               
               
                 PLRMLVETAQERNEPIFPALIYSSAMVWTVGMAKLDPSSQGALQLPYDPEMEDSYDSFGEPSYPEVFE 
               
               
                   
               
               
                 PPLTGYPGEELEEEEERGVKLGLGDFIFYSVLVGKAAATGSGDWNTTLACFVAILIGLCLTLLLLAVF 
               
               
                   
               
               
                 KKALPALPISITFGLIFYFSTDNLVRPFMDTLASHQLYI 
               
            
           
         
       
     
     To determine the percent identity of two amino acid sequences, or of two nucleic acid sequences, the sequences are aligned for optimal comparison purposes (e.g., gaps can be introduced in one or both of a first and a second amino acid or nucleic acid sequence for optimal alignment and non-homologous sequences can be disregarded for comparison purposes). The length of a reference sequence aligned for comparison purposes is at least 80% of the length of the reference sequence, and in some embodiments is at least 90% or 100%. The amino acid residues or nucleotides at corresponding amino acid positions or nucleotide positions are then compared. When a position in the first sequence is occupied by the same amino acid residue or nucleotide as the corresponding position in the second sequence, then the molecules are identical at that position. The percent identity between the two sequences is a function of the number of identical positions shared by the sequences, taking into account the number of gaps, and the length of each gap, which need to be introduced for optimal alignment of the two sequences. In another embodiment, the percent identity of two amino acid sequences can be assessed as a function of the conservation of amino acid residues within the same family of amino acids (e.g., positive charge, negative charge, polar and uncharged, hydrophobic) at corresponding positions in both amino acid sequences (e.g., the presence of an alanine residue in place of a valine residue at a specific position in both sequences shows a high level of conservation, but the presence of an arginine residue in place of an aspartate residue at a specific position in both sequences shows a low level of conservation). 
     For example, the percent identity between two amino acid sequences can be determined using the Needleman and Wunsch ((1970) J. Mol. Biol. 48:444-453) algorithm which has been incorporated into the GAP program in the GCG software package, using a Blossum scoring matrix, e.g., with default values for gap penalty, gap extend penalty of 4, and frameshift gap penalty. 
     Codon Optimized Presenilin 1 
     Codon optimization is desirable to express proteins in specific host cells—e.g., bacterial, mouse, human. It will be appreciated by those skilled in the art that as a result of the degeneracy of the genetic code, a multitude of cDNAs encoding human presenilin 1, some bearing minimal similarity to the cDNAs of any known and naturally occurring gene, may be produced. Thus, the invention contemplates each and every possible variation of cDNA that could be made by selecting combinations based on possible codon choices. These combinations are made in accordance with the standard triplet genetic code as applied to the polynucleotide encoding naturally occurring human presenilin variant, and all such variations are to be considered as being specifically disclosed. An exemplary codon-optimized human PSEN1 nucleotide sequence is disclosed herein—e.g., SEQ ID NO:9; See  FIG. 2 . This codon-optimized human PSEN1 nucleotide sequence was generated by substituting codons in the naturally occurring PSEN1 nucleotide sequence that occur at lower frequency in human cells for codons that occur at higher frequency in human cells. Codon-optimized human PSEN1 nucleotide sequences can include sequences wherein less than 100% of the codons are optimized, e.g., wherein only 20%, 30%, 40%, 50%, 60%, 70%, 80%, or 90% of the wild type non-optimized codons are optimized. The frequency of occurrence for codons can be computationally determined by methods known in the art. An exemplary calculation of these codon frequencies is disclosed in Table 1. 
     An exemplary codon-optimized human PSEN1 sequence is as follows: 
     
       
         
           
               
            
               
                 &gt;Codon optimized  Homo sapiens  presenilin 1 (PSEN1) 
               
               
                 cDNA 
               
               
                 (SEQ ID NO: 9) 
               
               
                 CGACGCCACCATGACAGAACTGCCTGCCCCCCTGAGCTACTTCCAGAACG 
               
               
                   
               
               
                 CCCAGATGAGCGAGGACAACCACCTGAGCAACACCGTGCGGAGCCAGAAC 
               
               
                   
               
               
                 GACAACAGAGAGCGGCAGGAACACAACGACAGGCGGAGCCTGGGACACCC 
               
               
                   
               
               
                 TGAGCCCCTGTCTAATGGCAGACCCCAGGGCAACAGCAGACAGGTGGTGG 
               
               
                   
               
               
                 AACAGGACGAGGAAGAGGACGAAGAACTGACCCTGAAGTACGGCGCCAAG 
               
               
                   
               
               
                 CACGTGATCATGCTGTTCGTGCCCGTGACCCTGTGCATGGTCGTGGTGGT 
               
               
                   
               
               
                 GGCCACAATCAAGAGCGTGTCCTTCTACACCCGGAAGGACGGCCAGCTGA 
               
               
                   
               
               
                 TCTACACCCCCTTCACCGAGGACACCGAGACAGTGGGACAGAGAGCCCTG 
               
               
                   
               
               
                 CACAGCATCCTGAACGCCGCCATCATGATCAGCGTGATCGTCGTGATGAC 
               
               
                   
               
               
                 CATCCTGCTGGTGGTGCTGTACAAGTACCGGTGCTACAAAGTGATCCACG 
               
               
                   
               
               
                 CCTGGCTGATCATCAGCAGCCTGCTGCTGCTGTTCTTCTTTAGCTTCATC 
               
               
                   
               
               
                 TACCTGGGCGAGGTGTTCAAGACCTACAACGTGGCCGTGGACTACATCAC 
               
               
                   
               
               
                 CGTGGCCCTGCTGATCTGGAACTTCGGCGTCGTGGGCATGATCTCCATCC 
               
               
                   
               
               
                 ACTGGAAGGGCCCCCTGAGACTGCAGCAGGCCTACCTGATTATGATCTCC 
               
               
                   
               
               
                 GCCCTGATGGCCCTGGTGTTCATCAAGTACCTGCCCGAGTGGACCGCTTG 
               
               
                   
               
               
                 GCTGATCCTGGCCGTGATCTCCGTGTACGACCTGGTGGCCGTGCTGTGCC 
               
               
                   
               
               
                 CTAAGGGACCTCTGCGGATGCTGGTGGAAACCGCCCAGGAACGGAACGAG 
               
               
                   
               
               
                 ACACTGTTCCCTGCCCTGATCTACTCCAGCACAATGGTGTGGCTCGTGAA 
               
               
                   
               
               
                 CATGGCCGAGGGCGATCCTGAGGCCCAGCGGAGAGTGTCCAAGAACTCCA 
               
               
                   
               
               
                 AGTACAACGCCGAGAGCACCGAGCGCGAGAGCCAGGATACAGTGGCCGAG 
               
               
                   
               
               
                 AATGACGACGGCGGCTTCAGCGAGGAATGGGAGGCCCAGAGAGATAGCCA 
               
               
                   
               
               
                 CCTGGGCCCTCACAGAAGCACCCCTGAATCTAGAGCCGCCGTGCAGGAAC 
               
               
                   
               
               
                 TGAGCAGCTCCATTCTGGCCGGCGAGGACCCCGAAGAAAGAGGCGTGAAA 
               
               
                   
               
               
                 CTGGGCCTGGGCGACTTCATCTTCTACAGCGTGCTCGTGGGCAAGGCCAG 
               
               
                   
               
               
                 CGCCACAGCTAGCGGCGACTGGAACACCACAATCGCCTGCTTCGTGGCCA 
               
               
                   
               
               
                 TCCTGATCGGCCTGTGTCTGACACTTCTGCTGCTGGCCATCTTCAAGAAG 
               
               
                   
               
               
                 GCCCTGCCCGCCCTGCCTATCAGCATCACCTTCGGCCTGGTGTTTTACTT 
               
               
                   
               
               
                 CGCCACCGACTACCTGGTGCAGCCCTTCATGGACCAGCTGGCCTTCCACC 
               
               
                   
               
               
                 AGTTCTACATCTGA 
               
            
           
         
       
     
     
       
         
           
               
             
               
                 TABLE 1 
               
             
            
               
                   
               
               
                 Codon Usage Frequency Table in Humans (Source:  
               
               
                 Gen Script ®, GenScript Codon Usage Frequency Table Tool) 
               
            
           
           
               
               
               
               
               
            
               
                   
                 Amino 
                   
                 Frequency/ 
                   
               
               
                 Triplet 
                 acid 
                 Fraction 
                 Thousand 
                 Number 
               
               
                   
               
            
           
           
               
               
               
               
               
            
               
                 TTT 
                 F 
                 0.45 
                 16.9 
                 336562 
               
               
                 TTC 
                 F 
                 0.55 
                 20.4 
                 406571 
               
               
                 TTA 
                 L 
                 0.07 
                 7.2 
                 143715 
               
               
                 TTG 
                 L 
                 0.13 
                 12.6 
                 249879 
               
               
                 TAT 
                 Y 
                 0.43 
                 12 
                 239268 
               
               
                 TAC 
                 Y 
                 0.57 
                 15.6 
                 310695 
               
               
                 TAA 
                 * 
                 0.28 
                 0.7 
                 14322 
               
               
                 TAG 
                 * 
                 0.2 
                 0.5 
                 10915 
               
               
                 CTT 
                 L 
                 0.13 
                 12.8 
                 253795 
               
               
                 CTC 
                 L 
                 0.2 
                 19.4 
                 386182 
               
               
                 CTA 
                 L 
                 0.07 
                 6.9 
                 138154 
               
               
                 CTG 
                 L 
                 0.41 
                 40.3 
                 800774 
               
               
                 CAT 
                 H 
                 0.41 
                 10.4 
                 207826 
               
               
                 CAC 
                 H 
                 0.59 
                 14.9 
                 297048 
               
               
                 CAA 
                 Q 
                 0.25 
                 11.8 
                 234785 
               
               
                 CAG 
                 Q 
                 0.75 
                 34.6 
                 688316 
               
               
                 ATT 
                 I 
                 0.36 
                 15.7 
                 313225 
               
               
                 ATC 
                 I 
                 0.48 
                 21.4 
                 426570 
               
               
                 ATA 
                 I 
                 0.16 
                 7.1 
                 140652 
               
               
                 ATG 
                 M 
                 1 
                 22.3 
                 443795 
               
               
                 AAT 
                 N 
                 0.46 
                 16.7 
                 331714 
               
               
                 AAC 
                 N 
                 0.54 
                 19.5 
                 387148 
               
               
                 AAA 
                 K 
                 0.42 
                 24 
                 476554 
               
               
                 AAG 
                 K 
                 0.58 
                 32.9 
                 654280 
               
               
                 GTT 
                 V 
                 0.18 
                 10.9 
                 216818 
               
               
                 GTC 
                 V 
                 0.24 
                 14.6 
                 290874 
               
               
                 GTA 
                 V 
                 0.11 
                 7 
                 139156 
               
               
                 GTG 
                 V 
                 0.47 
                 28.9 
                 575438 
               
               
                 GAT 
                 D 
                 0.46 
                 22.3 
                 443369 
               
               
                 GAC 
                 D 
                 0.54 
                 26 
                 517579 
               
               
                 GAA 
                 E 
                 0.42 
                 29 
                 577846 
               
               
                 GAG 
                 E 
                 0.58 
                 40.8 
                 810842 
               
               
                 TCT 
                 S 
                 0.18 
                 14.6 
                 291040 
               
               
                 TCC 
                 S 
                 0.22 
                 17.4 
                 346943 
               
               
                 TCA 
                 S 
                 0.15 
                 11.7 
                 233110 
               
               
                 TCG 
                 S 
                 0.06 
                 4.5 
                 89429 
               
               
                 TGT 
                 C 
                 0.45 
                 9.9 
                 197293 
               
               
                 TGC 
                 C 
                 0.55 
                 12.2 
                 243685 
               
               
                 TGA 
                 * 
                 0.52 
                 1.3 
                 25383 
               
               
                 TGG 
                 W 
                 1 
                 12.8 
                 255512 
               
               
                 CCT 
                 P 
                 0.28 
                 17.3 
                 343793 
               
               
                 CCC 
                 P 
                 0.33 
                 20 
                 397790 
               
               
                 CCA 
                 P 
                 0.27 
                 16.7 
                 331944 
               
               
                 CCG 
                 P 
                 0.11 
                 7 
                 139414 
               
               
                 CGT 
                 R 
                 0.08 
                 4.7 
                 93458 
               
               
                 CGC 
                 R 
                 0.19 
                 10.9 
                 217130 
               
               
                 CGA 
                 R 
                 0.11 
                 6.3 
                 126113 
               
               
                 CGG 
                 R 
                 0.21 
                 11.9 
                 235938 
               
               
                 ACT 
                 T 
                 0.24 
                 12.8 
                 255582 
               
               
                 ACC 
                 T 
                 0.36 
                 19.2 
                 382050 
               
               
                 ACA 
                 T 
                 0.28 
                 14.8 
                 294223 
               
               
                 ACG 
                 T 
                 0.12 
                 6.2 
                 123533 
               
               
                 AGT 
                 S 
                 0.15 
                 11.9 
                 237404 
               
               
                 AGC 
                 S 
                 0.24 
                 19.4 
                 385113 
               
               
                 AGA 
                 R 
                 0.2 
                 11.5 
                 228151 
               
               
                 AGG 
                 R 
                 0.2 
                 11.4 
                 227281 
               
               
                 GCT 
                 A 
                 0.26 
                 18.6 
                 370873 
               
               
                 GCC 
                 A 
                 0.4 
                 28.5 
                 567930 
               
               
                 GCA 
                 A 
                 0.23 
                 16 
                 317338 
               
               
                 GCG 
                 A 
                 0.11 
                 7.6 
                 150708 
               
               
                 GGT 
                 G 
                 0.16 
                 10.8 
                 215544 
               
               
                 GGC 
                 G 
                 0.34 
                 22.8 
                 453917 
               
               
                 GGA 
                 G 
                 0.25 
                 16.3 
                 325243 
               
               
                 GGG 
                 G 
                 0.25 
                 16.4 
                 326879 
               
               
                   
               
            
           
         
       
     
     Mutated Presenilin 1 
     In some embodiments, the PS1 protein contains a mutation. In some embodiments, the mutation is a conservative substitution. Such changes include substituting any of isoleucine (I), valine (V), and leucine (L) for any other of these hydrophobic amino acids; aspartic acid (D) for glutamic acid (E) and vice versa; glutamine (Q) for asparagine (N) and vice versa; and serine (S) for threonine (T) and vice versa. Other substitutions can also be considered conservative, depending on the environment of the particular amino acid and its role in the three-dimensional structure of the protein. For example, glycine (G) and alanine (A) can frequently be interchangeable, as can alanine (A) and valine (V). Methionine (M), which is relatively hydrophobic, can frequently be interchanged with leucine and isoleucine, and sometimes with valine. Lysine (K) and arginine (R) are frequently interchangeable in locations in which the significant feature of the amino acid residue is its charge and the differing pK&#39;s of these two amino acid residues are not significant. Still other changes can be considered “conservative” in particular environments (see, e.g. Table III of US20110201052; pages 13-15 “Biochemistry” 2nd ED. Stryer ed (Stanford University); Henikoff et al., PNAS 1992 Vol 89 10915-10919; Lei et al., J Biol Chem 1995 May 19; 270(20):11882-6). 
     In some embodiments, the methods include introducing one or more additional mutations into the human PS1 sequence (SEQ ID NOs:5 or 6). Thus, in some embodiments, the sequence can be at least 80%, 85%, 90%, 95%, or 99% identical to at least 60%, 70%, 80%, 90%, or 100% of a human PS1. 
     To determine the percent identity of two amino acid sequences, or of two nucleic acid sequences, the sequences are aligned for optimal comparison purposes (e.g., gaps can be introduced in one or both of a first and a second amino acid or nucleic acid sequence for optimal alignment and non-homologous sequences can be disregarded for comparison purposes). The length of a reference sequence aligned for comparison purposes is typically at least 80% of the length of the reference sequence, and in some embodiments is at least 90% or 100%. The amino acid residues or nucleotides at corresponding amino acid positions or nucleotide positions are then compared. When a position in the first sequence is occupied by the same amino acid residue or nucleotide as the corresponding position in the second sequence, then the molecules are identical at that position (as used herein amino acid or nucleic acid “identity” is equivalent to amino acid or nucleic acid “homology”). The percent identity between the two sequences is a function of the number of identical positions shared by the sequences, taking into account the number of gaps, and the length of each gap, which need to be introduced for optimal alignment of the two sequences. In another embodiment, the percent identity of two amino acid sequences can be assessed as a function of the conservation of amino acid residues within the same family of amino acids (e.g., positive charge, negative charge, polar and uncharged, hydrophobic) at corresponding positions in both amino acid sequences (e.g., the presence of an alanine residue in place of a valine residue at a specific position in both sequences shows a high level of conservation, but the presence of an arginine residue in place of an aspartate residue at a specific position in both sequences shows a low level of conservation). 
     For purposes of the present methods, the comparison of sequences and determination of percent identity between two sequences can be accomplished using a Blossum 62 scoring matrix with a gap penalty of 12, a gap extend penalty of 4, and a frameshift gap penalty of 5. 
     Delivery Vectors 
     Codon-optimized nucleic acids encoding a PS1 polypeptide or therapeutically active fragment thereof can be incorporated into a gene construct to be used as a part of a gene therapy protocol. For example, described herein are targeted expression vectors for in vivo delivery and expression of a codon-optimized polynucleotide that encodes a PS1 polypeptide or active fragment thereof in particular cell types, especially cerebral cortical neuronal cells. Expression constructs of such components can be administered in any effective carrier, e.g., any formulation or composition capable of effectively delivering the component gene to cells in vivo. Approaches include insertion of the gene in viral vectors, preferably adeno-associated virus. Viral vectors typically transduce cells directly. 
     Viral vectors capable of highly efficient transduction of CNS neurons may be employed, including any serotypes of rAAV (e.g., AAV1-AAV12) vectors, recombinant or chimeric AAV vectors, as well as lentivirus or other suitable viral vectors. In some embodiments, a codon-optimized polynucleotide encoding PS1 is operably linked to promoter suitable for expression in the CNS. For example, a neuron subtype-specific specific promoter, such as the alpha-calcium/calmodulin kinase 2A promoter may be used to target excitatory neurons. Alternatively, a pan neuronal promoter, such as the synapsin I promoter, may be used to drive PS1 expression. Other exemplary promoters include, but are not limited to, a cytomegalovirus (CMV) early enhancer/promoter; a hybrid CMV enhance/chicken (3-actin (CBA) promoter; a promoter comprising the CMV early enhancer element, the first exon and first intron of the chicken β-actin gene, and the splice acceptor of the rabbit (3-globin gene (commonly call the “CAG promoter”); or a 1.6-kb hybrid promoter composed of a CMV immediate-early enhancer and CBA intron 1/exon 1 (commonly called the CAGGS promoter; Niwa et al. Gene, 108:193-199 (1991)). The CAGGS promoter (Niwa et al., 1991) has been shown to provide ubiquitous and long-term expression in the brain (Klein et al., Exp. Neurol. 176:66-74 (2002)). One approach for in vivo introduction of nucleic acid into a cell is by use of a viral vector containing nucleic acid, e.g., a codon-optimized cDNA encoding a PS1. Among other things, infection of cells with a viral vector has the advantage that a large proportion of the targeted cells can receive the nucleic acid. Additionally, molecules encoded within the viral vector, e.g., by a cDNA contained in the viral vector, are expressed efficiently in cells that have taken up viral vector nucleic acid. 
     A viral vector system particularly useful for delivery of nucleic acids is the adeno-associated virus (AAV). Adeno-associated virus is a naturally occurring defective virus that requires another virus, such as an adenovirus or a herpes virus, as a helper virus for efficient replication and a productive life cycle. (For a review see Muzyczka et al., Curr. Topics in Micro and Immunol. 158:97-129 (1992)). AAV vectors efficiently transduce various cell types and can produce long-term expression of transgenes in vivo. Although AAV vector genomes can persist within cells as episomes, vector integration has been observed (see for example Deyle and Russell, Curr Opin Mol Ther. 2009 August; 11(4): 442-447; Asokan et al., Mol Ther. 2012 April; 20(4): 699-708; Flotte et al., Am. J. Respir. Cell. Mol. Biol. 7:349-356 (1992); Samulski et al., J. Virol. 63:3822-3828 (1989); and McLaughlin et al., J. Virol. 62:1963-1973 (1989)). AAV vectors, such as AAV2, have been extensively used for gene augmentation or replacement and have shown therapeutic efficacy in a range of animal models as well as in the clinic; see, e.g., Mingozzi and High, Nature Reviews Genetics 12, 341-355 (2011); Deyle and Russell, Curr Opin Mol Ther. 2009 August; 11(4): 442-447; Asokan et al., Mol Ther. 2012 April; 20(4): 699-708. AAV vectors containing as little as 300 base pairs of AAV can be packaged and can produce recombinant protein expression. Protocols for producing recombinant retroviruses and for infecting cells in vitro or in vivo with such viruses are known in the art, e.g, can be found in Ausubel, et al., eds., Current Protocols in Molecular Biology, Greene Publishing Associates, (1989), Sections 9.10-9.14, and other standard laboratory manuals. The use of AAV vectors to deliver constructs for expression in the brain has been described, e.g., in Iwata et al., Sci Rep. 2013; 3:1472; Hester et al., Curr Gene Ther. 2009 October; 9(5):428-33; Doll et al., Gene Therapy 1996, 3(5):437-447; and Foley et al., J Control Release. 2014 Dec. 28; 196:71-8. 
     Thus, in some embodiments, the codon-optimized PSENlencoding nucleic acid is present in a vector for gene therapy, such as an AAV vector. In some instances, the AAV vector is selected from the group consisting of AAV1, AAV2, AAV3, AAV4, AAV5, AAV6, AAV7, AAV8, AAV9, AAVrh10, AAV11, and AAV12. In preferred embodiments, the AAV is AAV9 or AAVrh10. 
     A vector as described herein can be a pseudotyped vector. Pseudotyping provides a mechanism for modulating a vector&#39;s target cell population. For instance, pseudotyped AAV vectors can be utilized in various methods described herein. Pseudotyped vectors are those that contain the genome of one vector, e.g., the genome of one AAV serotype, in the capsid of a second vector, e.g., a second AAV serotype. Methods of pseudotyping are well known in the art. For instance, a vector may be pseudotyped with envelope glycoproteins derived from Rhabdovirus vesicular stomatitis virus (VSV) serotypes (Indiana and Chandipura strains), rabies virus (e.g., various Evelyn-Rokitnicki-Abelseth ERA strains and challenge virus standard (CVS)), Lyssavirus Mokola virus, a rabies-related virus, vesicular stomatitis virus (VSV), Mokola virus (MV), lymphocytic choriomeningitis virus (LCMV), rabies virus glycoprotein (RV-G), glycoprotein B type (FuG-B), a variant of FuG-B (FuG-B2) or Moloney murine leukemia virus (MuLV). A virus may be pseudotyped for transduction of one or more neurons or groups of cells. 
     Without limitation, illustrative examples of pseudotyped vectors include recombinant AAV2/1, AAV2/2, AAV2/5, AAV2/6, AAV2/7, AAV2/8, AAV9, AAVrh10, AAV11, and AAV12 serotype vectors. It is known in the art that such vectors may be engineered to include a transgene encoding a human protein or other protein. In particular instances, the present disclosures can include a pseudotyped AAV9 or AAVrh10 viral vector including a nucleic acid as disclosed herein. See  Viral Vectors for Gene Therapy: Methods and Protocols , ed. Machida, Humana Press, 2003. 
     In some instances, a particular AAV serotype vector may be selected based upon the intended use, e.g., based upon the intended route of administration. 
     Various methods for application of AAV vector constructs in gene therapy are known in the art, including methods of modification, purification, and preparation for administration to human subjects (see, e.g., Viral Vectors for Gene Therapy: Methods and Protocols, ed. Machida, Humana Press, 2003). In addition, AAV based gene therapy targeted to cells of the CNS has been described (see, e.g., U.S. Pat. Nos. 6,180,613 and 6,503,888). High titer AAV preparations can be produced using techniques known in the art, e.g., as described in U.S. Pat. No. 5,658,776 
     A vector construct refers to a polynucleotide molecule including all or a portion of a viral genome and a transgene. In some instances, gene transfer can be mediated by a DNA viral vector, such as an adenovirus (Ad) or adeno-associated virus (AAV). Other vectors useful in methods of gene therapy are known in the art. For example, a construct as disclosed herein can include an alphavirus, herpesvirus, retrovirus, lentivirus, or vaccinia virus. 
     Adenoviruses are a relatively well characterized group of viruses, including over 50 serotypes (see, e.g., WO 95/27071, which is herein incorporated by reference). Adenoviruses are tractable through the application of techniques of molecular biology and may not require integration into the host cell genome. Recombinant Ad-derived vectors, including vectors that reduce the potential for recombination and generation of wild-type virus, have been constructed (see, e.g., international patent publications WO 95/00655 and WO 95/11984, which are herein incorporated by reference). Wild-type AAV has high infectivity and is capable of integrating into a host genome with a high degree of specificity (see, e.g. Hermonat and Muzyczka 1984 Proc. Natl. Acad. Sci., USA 81:6466-6470 and Lebkowski et al. 1988 Mol. Cell. Biol. 8:3988-3996). 
     Non-native regulatory sequences, gene control sequences, promoters, non-coding sequences, introns, or coding sequences can be included in a nucleic acid as disclosed herein. The inclusion of nucleic acid tags or signaling sequences, or nucleic acids encoding protein tags or protein signaling sequences, is further contemplated herein. Typically, the coding region is operably linked with one or more regulatory nucleic acid components. 
     A promoter included in a nucleic acid as disclosed herein can be a tissue- or cell type-specific promoter, a promoter specific to multiple tissues or cell types, an organ-specific promoter, a promoter specific to multiple organs, a systemic or ubiquitous promoter, or a nearly systemic or ubiquitous promoter. Promoters having stochastic expression, inducible expression, conditional expression, or otherwise discontinuous, inconstant, or unpredictable expression are also included within the scope of the present disclosure. A promoter can include any of the above characteristics or other promoter characteristics known in the art. 
     In clinical settings, the gene delivery systems for the therapeutic gene can be introduced into a subject by any of a number of methods, each of which is familiar in the art. For instance, a pharmaceutical preparation of the gene delivery system can be introduced systemically, e.g., by intravenous injection, and specific transduction of the protein in the target cells will occur predominantly from specificity of transfection, provided by the gene delivery vehicle, cell-type or tissue-type expression due to the transcriptional regulatory sequences controlling expression of the receptor gene, or a combination thereof. In other embodiments, initial delivery of the recombinant gene is more limited, with introduction into the subject being quite localized. For example, the gene delivery vehicle can be introduced by catheter (see U.S. Pat. No. 5,328,470) or by stereotactic injection, e.g., optionally into the cisterna magna, cerebral ventricles, lumbar intrathecal space, direct injection into hippocampus (e.g., Chen et al., PNAS USA 91: 3054-3057 (1994)). In preferred embodiments, delivery methods of Presenilin-expressing virus include intravenous, intrathecal, intracerebroventricular, intracisternal, and stereotactic intraparenchymal administration. 
     The methods can be further optimized via preclinical testing to achieve the best rescue of neurodegeneration, dementia, synaptic dysfunction and molecular alteration in presenilin conditional double knockout mice and presenilin-1 knockin mice expressing FAD mutations. 
     The pharmaceutical preparation of the gene therapy construct can consist essentially of the gene delivery system in an acceptable diluent, or can comprise a slow release matrix in which the gene delivery vehicle is embedded. Alternatively, where the complete gene delivery system can be produced intact from recombinant cells, e.g., retroviral vectors, the pharmaceutical preparation can comprise one or more cells, which produce the gene delivery system. 
     Delivery Formulations and Pharmaceutical Compositions 
     In some embodiments, polynucleotides as disclosed herein for delivery to a target tissue in vivo are encapsulated or associated with in a nanoparticle. Methods for nanoparticle packaging are well known in the art, and are described, for example, in Bose S, et al (Role of Nucleolin in Human Parainfluenza Virus Type 3 Infection of Human Lung Epithelial Cells. J. Virol. 78:8146. 2004); Dong Y et al. Poly(d,l-lactide-co-glycolide)/montmorillonite nanoparticles for oral delivery of anticancer drugs. Biomaterials 26:6068. 2005); Lobenberg R. et al (Improved body distribution of 14C-labelled AZT bound to nanoparticles in rats determined by radioluminography. J Drug Target 5:171.1998); Sakuma S R et al (Mucoadhesion of polystyrene nanoparticles having surface hydrophilic polymeric chains in the gastrointestinal tract. Int J Pharm 177:161. 1999); Virovic L et al. Novel delivery methods for treatment of viral hepatitis: an update. Expert Opin Drug Deliv 2:707.2005); and Zimmermann E et al, Electrolyte- and pH-stabilities of aqueous solid lipid nanoparticle (SLN) dispersions in artificial gastrointestinal media. Eur J Pharm Biopharm 52:203. 2001). In some embodiments, one or more polynucleotides is delivered to a target tissue in vivo in a vesicle, e.g. a liposome (see Langer, Science 249:1527-1533 (1990); Treat et al., in Liposomes in the Therapy of Infectious Disease and Cancer, Lopez-Berestein and Fidler (eds.), Liss, New York, pp. 353-365 (1989); Lopez-Berestein, ibid., pp. 317-327; see generally ibid). In some embodiments, lipid-based nanoparticles (LNP) are used; see, e.g., Robinson et al., Mol Ther. 2018 Aug. 1; 26(8):2034-2046; U.S. Pat. No. 9,956,271B2. 
     The present methods and compositions can include microvesicles or a preparation thereof, that contains one or more therapeutic molecules—e.g., polynucleotides or RNA—described herein. “Microvesicles”, as the term is used herein, refers to membrane-derived microvesicles, which includes a range of extracellular vesicles, including exosomes, microparticles and shed microvesicles secreted by many cell types under both normal physiological and pathological conditions. See, e.g., EP2010663B1. The methods and compositions described herein can be applied to microvesicles of all sizes; in one embodiment, 30 to 200 nm, in one embodiment, 30 to 800 nm, in one embodiment, up to 2 um. The methods and compositions described herein can also be more broadly applied to all extracellular vesicles, a term which encompasses exosomes, shed microvesicles, oncosomes, ectosomes, and retroviral-like particles. Such a microvesicle or preparation is produced by the herein described methods. As the term is used herein, a microvesicle preparation refers to a population of microvesicles obtained/prepared from the same cellular source. Such a preparation is generated, for example, in vitro, by culturing cells expressing the nucleic acid molecule of the instant invention and isolating microvesicles produced by the cells. Methods of isolating such microvesicles are known in the art (Thery et al., Isolation and characterization of exosomes from cell culture supernatants and biological fluids, in Current Protocols Cell Biology, Chapter 3, 322, (John Wiley, 2006); Palmisano et al., (Mol Cell Proteomics. 2012 August; 11(8):230-43) and Waldenström et al., ((2012) PLoS ONE 7(4): e34653.doi: 10.1371/journal.pone.0034653)), some examples of which are described herein. Such techniques for isolating microvesicles from cells in culture include, without limitation, sucrose gradient purification/separation and differential centrifugation, and can be adapted for use in a method or composition described herein. See, e.g., EP2010663B1. 
     In some embodiments, the microvesicles are isolated by gentle centrifugation (e.g., at about 300 g) of the culture medium of the donor cells for a period of time adequate to separate cells from the medium (e.g., about 15 minutes). This leaves the microvesicles in the supernatant, to thereby yield the microvesicle preparation. In one embodiment, the culture medium or the supernatant from the gentle centrifugation, is more strongly centrifuged (e.g., at about 16,000 g) for a period of time adequate to precipitate cellular debris (e.g., about 30 minutes). This leaves the microvesicles in the supernatant, to thereby yield the microvesicle preparation. In one embodiment, the culture medium, the gentle centrifuged preparation, or the strongly centrifuged preparation is subjected to filtration (e.g., through a 0.22 um filter or a 0.8 um filter, whereby the microvesicles pass through the filter. In one embodiment, the filtrate is subjected to a final ultracentrifugation (e.g. at about 110,000 g) for a period of time that will adequately precipitate the microvesicles (e.g. for about 80 minutes). The resulting pellet contains the microvesicles and can be resuspended in a volume of buffer that yields a useful concentration for further use, to thereby yield the microvesicle preparation. In one embodiment, the microvesicle preparation is produced by sucrose density gradient purification. In one embodiment, the microvesicles are further treated with DNAse (e.g., DNAse I) and/or RNAse and/or proteinase to eliminate any contaminating DNA, RNA, or protein, respectively, from the exterior. In one embodiment, the microvesicle preparation contains one or more RNAse inhibitors. 
     The molecules contained within the microvesicle preparation will comprise the therapeutic molecule. Typically the microvesicles in a preparation will be a heterogeneous population, and each microvesicle will contain a complement of molecule that may or may not differ from that of other microvesicles in the preparation. The content of the therapeutic molecules in a microvesicle preparation can be expressed either quantitatively or qualitatively. One such method is to express the content as the percentage of total molecules within the microvesicle preparation. By way of example, if the therapeutic molecule is an mRNA, the content can be expressed as the percentage of total RNA content, or alternatively as the percentage of total mRNA content, of the microvesicle preparation. Similarly, if the therapeutic molecule is a protein, the content can be expressed as the percentage of total protein within the microvesicles. In one embodiment, therapeutic microvesicles, or a preparation thereof, produced by the method described herein contain a detectable, statistically significantly increased amount of the therapeutic molecule as compared to microvesicles obtained from control cells (cells obtained from the same source which have not undergone scientific manipulation to increase expression of the therapeutic molecule). In one embodiment, the therapeutic molecule is present in an amount that is at least about 10%, 20%, 30% 40%, 50%, 60%, 70% 80% or 90%, more than in microvesicles obtained from control cells. Higher levels of enrichment may also be achieved. In one embodiment, the therapeutic molecule is present in the microvesicle or preparation thereof, at least 2 fold more than control cell microvesicles. Higher fold enrichment may also be obtained (e.g., 3, 4, 5, 6, 7, 8, 9 or 10 fold). 
     In one embodiment, a relatively high percentage of the microvesicle content is the therapeutic molecule (e.g., achieved through overexpression or specific targeting of the molecule to microvesicles). In one embodiment, the microvesicle content of the therapeutic molecule is at least about 10%, 20%, 30% 40%, 50%, 60%, 70% 80% or 90%, of the total (like) molecule content (e.g., the therapeutic molecule is an mRNA and is about 10% of the total mRNA content of the microvesicle). Higher levels of enrichment may also be achieved. In one embodiment, the therapeutic molecule is present in the microvesicle or preparation thereof, at least 2 fold more than all other such (like) molecules. Higher fold enrichment may also be obtained (e.g., 3, 4, 5, 6, 7, 8, 9 or 10 fold). 
     EXAMPLES 
     The invention is further described in the following examples, which do not limit the scope of the invention described in the claims. 
     Example 1: Dose-Dependent Rescue of γ-Secretase Activity in MEFs with Varying PS Genotypes: PS1 +/+ , PS1 L435F/+ , PS1 L435F/L435F , PS1 −/−  and PS1 −/− ; PS2 −/−   
     To determine whether reduced γ-secretase activity associated with PSEN1 mutations can be corrected by introduction of wild-type (WT) hPS1, primary MEFs from embryos carrying varying PS genotypes, PS1 +/+ , PS1 L435F/+ , PS1 +/− , PS1 L435F/L435F , PS1 −/−  and PS1 −/− ; PS2 −/−  (DKO) were derived. The immortalized MEFs were transiently transfected with CMV-N ΔE, and γ-secretase activity was evaluated by measuring the levels of NICD and PS1 NTF/CTF. The NICD levels were reduced in a PS1 dosage sensitive manner, and were undetectable in DKO cells ( FIG. 1A ). The NICD levels were reduced but detectable in PS1 L435F/L435F  MEFs (“L435F KI/KI” MEFs) and PS1 −/−  MEFs ( FIG. 1A ), whereas de novo NICD production was undetectable by in vitro γ-secretase assay using L435F KI/KI and PS1 −/−  embryonic brains (Xia et al., Neuron. 2015 Mar. 4; 85(5):967-81). Without wishing to be bound to a particular theory, applicant submits that this may be due to lower levels of PS2 normally expressed in the embryonic brain relative to MEFs, leading to lower overall PS activity in L435F KI/KI and PS1 −/−  brains, relative to MEFs. To test this hypothesis, the γ-secretase activity was measured in PS1 L435F/+ ; PS2 −/−  MEFs and compared to PS1 L435F/+  MEFs. The γ-secretase activity was lower in PS1 L435F/+ ; PS2 −/−  MEFs compared to PS1 L435F/+  MEFs ( FIG. 1B ). 
     To determine whether the impaired γ-secretase activity in various PS mutant MEFs can be rescued by introduction of WT hPS1, varying amounts (0, 20, 40, 80 ng) of wild-type hPS1 cDNA (pCI-hPS1) were transfected into MEFs, along with CMV-N ΔE. Notably, increasing amounts of pC1-hPS1 transfected into the MEFs resulted in accumulation of PS1 protein and restored levels of PS1 NTF and NICD in mutant (PS1 L435F/+ , PS1 L435F/+ ; PS2 −/−  and DKO) MEFs ( FIG. 1B ). These results indicate that exogenous WT hPS1 can rescue the impaired γ-secretase activity in in various PS mutant MEFs. 
     Example 2: Development of an Optimized Wild-Type Human PS1 In Vitro Expression System 
     Methods 
     Cell Culture and Transfection 
     P sen-null mouse embryo fibroblasts (MEFs) lacking endogenous PS1 and PS2 were maintained in DMEM supplemented with 10% FBS and transiently transfected with plasmids expressing either the wild-type endogenous hPSEN1 cDNA (wt_PS1) or the codon optimized hPSEN1 cDNA (opti_PS1), with or without the γ-secretase reporter, CMV-N ΔE, using Lipofectamine 3000 according to instruction. Cell lysates were collected at 24 h time point. 
     Western Blotting 
     Cell lysates were subjected to SDS-PAGE, and proteins were transferred to nitrocellulose membranes. After blocking in TBST/5% non-fat dry milk, membranes were incubated overnight with primary antibodies. To control for loading, membranes were stripped and re-probed with anti-β-actin antibody. Band intensity was quantified using ImageJ software, and results were normalized to β-actin levels. Antibodies used included rat anti-PS1-NTF, rabbit anti-cleaved Notch (Val1744) (NICD), and mouse anti-β-actin. 
     Results 
     First, we expressed wt_hPS1 or opti_hPS1 in Psen-null MEFs to eliminate any contribution of endogenous PS1 or PS2. We detected a progressive increase in the levels of PS1 NTF when Psen-null MEFs were transfected with increasing amounts of vector encoding wt_hPS1 or opti_hPS1. Optimized hPS1 consistently expressed more PS1 relative to wt_hPS1 ( FIGS. 3A-3B ). 
     NotchΔE as substrate for γ-secretase-mediated cleavage was co-expressed with wt_hPS1 or opti_hPS1 to measure γ-secretase activity directly. Notch is cleaved by γ-secretase to release the Notch intracellular domain (NICD). We evaluated the dose-response relationships for γ-secretase-mediated cleavage of NotchΔE to produce NICD as a function of γ-secretase activity. Production of NICD increased linearly across the lower amounts of PS1 vectors transfected, but close to saturation at higher levels. More importantly, for each point, optimized hPS1 has higher γ-secretase activity ( FIG. 3 ). 
     Secreted endogenous Aβ40 and 42 levels are measured in the medium using ELISA. Another γ-secretase substrate, APP C99, is used to evaluate γ-secretase activity by transiently transfecting CMV-C99 into each of the MEF cell lines, along with increasing amounts of pCI-hPS1opti. Primary and immortalized C410Y, E280A and D385A KI/+ and KUKI MEFs as well as PS1 KI/+; PS2−/− MEFs are established. Whether KI/+ and KUKI MEFs recapitulate similar phenotypes as KI/+ and KUKI brains is determined, and whether introduction of WT hPS1 can restore the reduced γ-secretase activity in a dose-dependent manner is measured by NICD and AICD production. The experiment is repeated with multiple independent MEF cell lines per genotype. Power analysis is performed to determine the sample size and the number of independent experiments needed to complete the study. 
     Example 3: Development of an Optimized Wild-Type Human PS1 In Vivo Expression System 
     Transgenic mice are developed that express human PSEN1 wild-type cDNA constitutively or inducibly under the control of the CAMK2A promoter. To maximize PS1 production and activity, hPS1 was codon-optimized (hPS1opti), and then PS1 levels and γ-secretase activity were compared between the endogenous hPS1 and hPS1opti cDNA by co-transfecting increasing amounts of either pCI-hPS1 or pCI-hPS1opti and CMV-NΔE into PS DKO MEFs. Relative to the endogenous hPS1 cDNA, the hPS1opti cDNA resulted in higher levels of PS1 NTF and higher γ-secretase activity, measured by NICD production ( FIG. 3 ). 
     Example 4: Determine Whether Postnatal Delivery of hPS1opti can Rescue Phenotypes in PS cDKO Mice 
     The vectors listed in Table 2 have been prepared. The vector that confers the highest GFP staining at 4 and 8 weeks after injection is identified. To do this, PS cDKO pups (lots−10 breeding cages of F/F; −/−; Cre/Cre crossed with F/F; −/−) are injected at P0-2. 
     
       
         
           
               
             
               
                 TABLE 2 
               
             
            
               
                   
               
               
                 List of vectors 
               
            
           
           
               
               
            
               
                 UID 
                 Description 
               
               
                   
               
            
           
           
               
               
            
               
                 01 
                 AAV9/mCaMKII-intron-hPS1opti-T2A-EGFP-SV40pA 
               
               
                 02 
                 AAV9/hCaMKII-intron-hPS1opti-T2A-EGFP-SV40pA 
               
               
                 03 
                 AAV9/hCaMKII-intron-EGFP-SV40pA 
               
               
                 04 
                 AAV9/hSynI-intron-hPS1opti-T2A-EGFP-SV40pA 
               
               
                 05 
                 AAV9/hCaMKII-intron-hPS1opti-SV40pA 
               
               
                 06 
                 AAV9/hSynI-intron-hPS1opti-SV40A 
               
               
                   
               
            
           
         
       
     
     An AAV—e.g., AAV9/hCaMKII-intron-hPS1opti-T2A-EGFP-SV40 pA or AAV9/hCaMKII-intron-EGFP-SV40 pA—is selected and injected into PS cDKO mice at P0-P2 by ICV. Western analysis is performed at 4 and 8 weeks of age to determine levels of PS1, APP, Nicastrin, PEN-2; control and PS cDKO mice are included at 4 and 8 weeks as additional controls. Gamma secretase activity, NICD production, and Aβ levels (by ELISA of cortical lysates) are measured at 8 weeks of age. Additionally, at 2 months of age electrophysiological analysis—e.g., Schaffer collateral, PPF, FF, and LTP—is performed, and behavioral analysis is performed at 2-3 months of age—e.g., watermaze. Neuropath analysis is performed at 6 months of age. 
     Example 5: Determine Whether Postnatal Delivery of hPS1opti can Rescue Phenotypes Caused by FAD Mutations 
     Mice listed in Table 3 are generated, and analyzed using western blot analysis to measure PS1 and APP levels; in vitro gamma-secretase activity assay; ELISA, electrophysiological analysis at 6 months of age; behavior analysis at 6 and 12 months of age; a neuropath analysis at 6, 12, and 18 months of age.