Patent Publication Number: US-2002013458-A1

Title: Enzymatic nucleic acid treatment of disases or conditions related to hepatitis c virus infection

Description:
[0001] This patent application is a continuation-in-part of Blatt et al., U.S. Ser. No. 09/274,553, filed Mar. 22, 1999 and Blatt et al., U.S. Ser. No. 09/257,608, filed Feb. 24, 1999, which both claim the benefit of Blatt et al., U.S. Ser. No. 60/100,842, filed Sep. 18, 1998, and McSwiggen et al., U.S. Ser. No. 60/083,217 filed Apr. 27, 1998, all of these earlier applications are entitled “ENZYMATIC NUCLEIC ACID TREATMENT OF DISEASES OR CONDITIONS RELATED TO HEPATITIS C VIRUS INFECTION”. Each of these applications are hereby incorporated by reference herein in their entirety including the drawings. 
    
    
     
       BACKGROUND OF THE INVENTION  
       [0002] This invention relates to methods and reagents for the treatment of diseases or conditions relating to the hepatitis C virus (HCV) infection.  
       [0003] The following is a discussion of relevant art, none of which is admitted to be prior art to the present invention.  
       [0004] In 1989, the HCV was determined to be an RNA virus and was identified as the causative agent of most non-A non-B viral Hepatitis (Choo et al.,  Science.  1989; 244:359-362). Unlike retroviruses such as HIV, HCV does not go though a DNA replication phase and no integrated forms of the viral genome into the host chromosome have been detected (Houghton et al.,  Hepatology  1991;14:381-388). Rather, replication of the coding (plus) strand is mediated by the production of a replicative (minus) strand leading to the generation of several copies of plus strand HCV RNA. The genome consists of a single, large, open-reading frame that is translated into a polyprotein (Kato et al.,  FEBS Letters.  1991; 280: 325-328). This polyprotein subsequently undergoes post-translational cleavage, producing several viral proteins (Leinbach et al.,  Virology.  1994: 204:163-169).  
       [0005] Examination of the 9.5-kilobase genome of HCV has demonstrated that the viral nucleic acid can mutate at a high rate (Smith et al.,  Mol. Evol.  1997 45:238-246). This rate of mutation has led to the evolution of several distinct genotypes of HCV that share approximately 70% sequence identity (Simmonds et al.,  J. Gen. Virol  1994; 75:1053-1061). It is important to note that these sequences are evolutionarily quite distant. For example, the genetic identity between humans and primates such as the chimpanzee is approximately 98%. In addition, it has been demonstrated that an HCV infection in an individual patient is composed of several distinct and evolving quasi-species that have 98% identity at the RNA level. Thus, the HCV genome is hypervariable and continuously changing. Although the HCV genome is hypervariable, there are 3 regions of the genome that are highly conserved. These conserved sequences occur in the 5′ and 3′ non-coding regions as well as the 5′-end of the core protein coding region and are thought to be vital for HCV RNA replication as well as translation of the HCV polyprotein. Thus, therapeutic agents that target these conserved HCV genomic regions may have a significant impact over a wide range of HCV genotypes. Moreover, it is unlikely that drug resistance will occur with ribozymes specific to conserved regions of the HCV genome. In contrast, therapeutic modalities that target inhibition of enzymes such as the viral proteases or helicase are likely to result in the selection for drug resistant strains since the RNA for these viral encoded enzymes is located in the hypervariable portion of the HCV genome.  
       [0006] After initial exposure to HCV, the patient will experience a transient rise in liver enzymes, which indicates that inflammatory processes are occurring (Alter et al., IN: Seeff L B, Lewis J H, eds.  Current Perspectives in Hepatology.  New York: Plenum Medical Book Co; 1989:83-89). This elevation in liver enzymes will occur at least 4 weeks after the initial exposure and may last for up to two months (Farci et al.,  New England Journal of medicine.  1991:325:98-104). Prior to the rise in liver enzymes, it is possible to detect HCV RNA in the patient&#39;s serum using RT-PCR analysis (Takahashi et al.,  American Journal of Gastroenterology.  1993:88:2:240-243). This stage of the disease is called the acute stage and usually goes undetected since 75% of patients with acute viral hepatitis from HCV infection are asymptomatic. The remaining 25% of these patients develop jaundice or other symptoms of hepatitis.  
       [0007] Acute HCV infection is a benign disease, however, and as many as 80% of acute HCV patients progress to chronic liver disease as evidenced by persistent elevation of serum alanine aminotransferase (ALT) levels and by continual presence of circulating HCV RNA (Sherlock,  Lancet  1992; 339:802). The natural progression of chronic HCV infection over a 10 to 20 year period leads to cirrhosis in 20 to 50% of patients (Davis et al.,  Infectious Agents and Disease  1993;2:150:154) and progression of HCV infection to hepatocellular carcinoma has been well documented (Liang et al.,  Hepatology.  1993; 18:1326-1333; Tong et al,  Western Journal of Medicine,  1994; Vol. 160, No. 2: 133-138). There have been no studies that have determined sub-populations that are most likely to progress to cirrhosis and/or hepatocellular carcinoma, thus all patients have an equal risk of progression.  
       [0008] It is important to note that the survival for patients diagnosed with hepatocellular carcinoma is only 0.9 to 12.8 months from initial diagnosis (Takahashi et aL,  American Journal of Gastroenterology.  1993:88:2:240-243). Treatment of hepatocellular carcinoma with chemotherapeutic agents has not proven effective and only 10% of patients will benefit from surgery due to extensive tumor invasion of the liver (Trinchet et al.,  Presse Medicine.  1994:23:831-833). Given the aggressive nature of primary hepatocellular carcinoma, the only viable treatment alternative to surgery is liver transplantation (Pichlmayr et al.,  Hepatology.  1994:20:33S-40S).  
       [0009] Upon progression to cirrhosis, patients with chronic HCV infection present with clinical features, which are common to clinical cirrhosis regardless of the initial cause (D&#39;Amico et al.,  Digestive Diseases and Sciences.  1986;31:5: 468-475). These clinical features may include: bleeding esophageal varices, ascites, jaundice, and encephalopathy (Zakim D, Boyer TD.  Hepatology  a textbook of liver disease. Second Edition Volume 1. 1990 W. B. Saunders Company. Philadelphia). In the early stages of cirrhosis, patients are classified as compensated meaning that although liver tissue damage has occurred, the patient&#39;s liver is still able to detoxify metabolites in the bloodstream. In addition, most patients with compensated liver disease are asymptomatic and the minority with symptoms report only minor symptoms such as dyspepsia and weakness. In the later stages of cirrhosis, patients are classified as decompensated meaning that their ability to detoxify metabolites in the bloodstream is diminished and it is at this stage that the clinical features described above will present.  
       [0010] In 1986, D&#39;Amico et al. described the clinical manifestations and survival rates in 1155 patients with both alcoholic and viral associated cirrhosis (D&#39;Amico supra). Of the 1155 patients, 435 (37%) had compensated disease although 70% were asymptomatic at the beginning of the study. The remaining 720 patients (63%) had decompensated liver disease with 78% presenting with a history of ascites, 31% with jaundice, 17% had bleeding and 16% had encephalopathy. Hepatocellular carcinoma was observed in six (0.5%) patients with compensated disease and in 30 (2.6%) patients with decompensated disease.  
       [0011] Over the course of six years, the patients with compensated cirrhosis developed clinical features of decompensated disease at a rate of 10% per year. In most cases, ascites was the first presentation of decompensation. In addition, hepatocellular carcinoma developed in 59 patients who initially presented with compensated disease by the end of the six-year study.  
       [0012] With respect to survival, the D&#39;Amico study indicated that the five-year survival rate for all patients on the study was only 40%. The six-year survival rate for the patients who initially had compensated cirrhosis was 54% while the six-year survival rate for patients who initially presented with decompensated disease was only 21%. There were no significant differences in the survival rates between the patients who had alcoholic cirrhosis and the patients with viral related cirrhosis. The major causes of death for the patients in the D&#39;Amico study were liver failure in 49%; hepatocellular carcinoma in 22%; and, bleeding in 13% (D&#39;Amico supra).  
       [0013] Chronic Hepatitis C is a slowly progressing inflammatory disease of the liver, mediated by a virus (HCV) that can lead to cirrhosis, liver failure and/or hepatocellular carcinoma over a period of 10 to 20 years. In the US, it is estimated that infection with HCV accounts for 50,000 new cases of acute hepatitis in the United States each year (NIH Consensus Development Conference Statement on Management of Hepatitis C March 1997). The prevalence of HCV in the United States is estimated at 1.8% and the CDC places the number of chronically infected Americans at approximately 4.5 million people. The CDC also estimates that up to 10,000 deaths per year are caused by chronic HCV infection. The prevalence of HCV in the United States is estimated at 1.8% and the CDC places the number of chronically infected Americans at approximately 4.5 million people. The CDC also estimates that up to 10,000 deaths per year are caused by chronic HCV infection.  
       [0014] Numerous well controlled clinical trials using interferon (IFN-alpha) in the treatment of chronic HCV infection have demonstrated that treatment three times a week results in lowering of serum ALT values in approximately 50% (range 40% to 70%) of patients by the end of 6 months of therapy (Davis et al.,  New England Journal of Medicine  1989; 321:1501-1506; Marcellin et al.,  Hepatology.  1991; 13:393-397; Tong et al.,  Hepatology  1997:26:747-754; Tong et al.,  Hepatology  1997 26(6): 1640-1645). However, following cessation of interferon treatment, approximately 50% of the responding patients relapsed, resulting in a “durable” response rate as assessed by normalization of serum ALT concentrations of approximately 20 to 25%.  
       [0015] In recent years, direct measurement of the HCV RNA has become possible through use of either the branched-DNA or Reverse Transcriptase Polymerase Chain Reaction (RT-PCR) analysis. In general, the RT-PCR methodology is more sensitive and leads to more accurate assessment of the clinical course (Tong et al., supra). Studies that have examined six months of type 1 interferon therapy using changes in HCV RNA values as a clinical endpoint have demonstrated that up to 35% of patients will have a loss of HCV RNA by the end of therapy (Marcellin et al., supra). However, as with the ALT endpoint, about 50% of the patients relapse six months following cessation of therapy resulting in a durable virologic response of only 12% (Marcellin et al., supra). Studies that have examined 48 weeks of therapy have demonstrated that the sustained virological response is up to 25% (NIH consensus statement: 1997). Thus, standard of care for treatment of chronic HCV infection with type 1 interferon is now 48 weeks of therapy using changes in HCV RNA concentrations as the primary assessment of efficacy (Hooftiagle et al.,  New England Journal of Medicine  1997; 336(5) 347-356).  
       [0016] Side effects resulting from treatment with type 1 interferons can be divided into four general categories, which include 1. Influenza-like symptoms; 2. Neuropsychiatric; 3. Laboratory abnormalities; and, 4. Miscellaneous (Dusheiko et al.,  Journal of Viral Hepatitis,  1994:1:3-5). Examples of influenza-like symptoms include: fatigue, fever; myalgia; malaise; appetite loss; tachycardia; rigors; headache and arthralgias. The influenza-like symptoms are usually short-lived and tend to abate after the first four weeks of dosing (Dushieko et al., supra). Neuropsychiatric side effects include: irritability, apathy; mood changes; insomnia; cognitive changes and depression. The most important of these neuropsychiatric side effects is depression and patients who have a history of depression should not be given type 1 interferon. Laboratory abnormalities include; reduction in myeloid cells including granulocytes, platelets and to a lesser extent red blood cells. These changes in blood cell counts rarely lead to any significant clinical sequellae (Dushieko et al., supra). In addition, increases in triglyceride concentrations and elevations in serum alanine and aspartate aminotransferase concentration have been observed. Finally, thyroid abnormalities have been reported. These thyroid abnormalities are usually reversible after cessation of interferon therapy and can be controlled with appropriate medication while on therapy. Miscellaneous side effects include nausea; diarrhea; abdominal and back pain; pruritus; alopecia; and rhinorrhea. In general, most side effects will abate after 4 to 8 weeks of therapy (Dushieko et aL, supra).  
       [0017] Welch et al.,  Gene Therapy  1996 3(11): 994-1001 describe in vitro an in vivo studies with two vector expressed hairpin ribozymes targeted against hepatitis C virus.  
       [0018] Sakamoto et al.,  J. Clinical Investigation  1996 98(12): 2720-2728 describe intracellular cleavage of hepatitis C virus RNA and inhibition of viral protein translation by certain vector expressed hammerhead ribozymes.  
       [0019] Lieber et al.,  J. Virology  1996 70(12): 8782-8791 describe elimination of hepatitis C virus RNA in infected human hepatocytes by adenovirus-mediated expression of certain hammerhead ribozymes.  
       [0020] Ohkawa et al., 1997,  J. Hepatology,  27; 78-84, describe in vitro cleavage of HCV RNA and inhibition of viral protein translation using certain in vitro transcribed hammerhead ribozymes.  
       [0021] Barber et al., International PCT Publication No. WO 97/32018, describe the use of an adenovirus vector to express certain anti-hepatitis C virus hairpin ribozymes.  
       [0022] Kay et al., International PCT Publication No. WO 96/18419, describe certain recombinant adenovirus vectors to express anti-HCV hammerhead ribozyme.  
       [0023] Yamada et al., Japanese Patent Application No. JP 07231784 describe a specific poly-(L)-lysine conjugated hammerhead ribozyme targeted against HCV.  
       [0024] Draper, U.S. Pat. Nos. 5,610,054 and 5,869,253, describe enzymatic nucleic acid molecules capable of inhibiting replication of HCV.  
       SUMMARY OF THE INVENTION  
       [0025] This invention relates to ribozymes, or enzymatic nucleic acid molecules, directed to cleave RNA species of hepatitis C virus (HCV) and/or encoded by the HCV. In particular, applicant describes the selection and function of ribozymes capable of specifically cleaving HCV RNA. Such ribozymes may be used to treat diseases associated with HCV infection.  
       [0026] Due to the high sequence variability of the HCV genome, selection of ribozymes for broad therapeutic applications would likely involve the conserved regions of the HCV genome. Specifically, the present invention describes hammerhead ribozymes that would cleave in the conserved regions of the HCV genome. A list of the thirty hammerhead ribozymes derived from the conserved regions (5′-Non Coding Region (NCR), 5′-end of core protein coding region, and 3′-NCR) of the HCV genome is shown in Table IV. In general, Applicant has found that enzymatic nucleic acid molecules that cleave sites located in the 5′ end of the HCV genome would block translation while ribozymes that cleave sites located in the 3′ end of the genome would block RNA replication. Approximately 50 HCV isolates have been identified and a sequence alignment of these isolates from genotypes 1a, 1b, 2a, 2b, 2c, 3a, 3b, 4a, 5a, and 6 was performed. These alignments were used by the Applicant to identify 30 hammerhead ribozymes sites within regions highly conserved between genotypes. Twenty-three ribozyme sites were identified in regions of greatest homology within the conserved region. Therefore, one ribozyme can be designed to cleave all the different isolates of HCV. According to the Applicant, ribozymes designed against conserved regions of various HCV isolates will enable efficient inhibition of HCV replication in diverse patient populations and may ensure the effectiveness of the ribozymes against HCV quasi species which evolve due to mutations in the non-conserved regions of the HCV genome.  
       [0027] In another preferred embodiment, the invention features the use of an enzymatic nucleic acid molecule, preferably in the hammerhead, NCH motif (Inozyme), G-cleaver, amberzyme, zinzyme and/or DNAzyme motif, to inhibit the expression of HCV RNA.  
       [0028] In yet another preferred embodiment, the invention features the use of an enzymatic nucleic acid molecule, preferably in the hammerhead, Inozyme, G-cleaver, amberzyme, zinzyme and/or DNAzyme motif, to inhibit the expression of HCV minus strand RNA.  
       [0029] By “inhibit” it is meant that the activity of HCV or level of RNAs or equivalent RNAs encoding one or more protein subunits of HCV is reduced below that observed in the absence of the nucleic acid molecules of the invention. In one embodiment, inhibition with enzymatic nucleic acid molecule preferably is below that level observed in the presence of an enzymatically inactive or attenuated molecule that is able to bind to the same site on the target RNA, but is unable to cleave that RNA. In another embodiment, inhibition of HCV genes with the nucleic acid molecule of the instant invention is greater than in the presence of the nucleic acid molecule than in its absence.  
       [0030] By “enzymatic nucleic acid molecule” it is meant a nucleic acid molecule which has complementarity in a substrate binding region to a specified gene target, and also has an enzymatic activity which is active to specifically cleave target RNA. That is, the enzymatic nucleic acid molecule is able to intermolecularly cleave RNA and thereby inactivate a target RNA molecule. These complementary regions allow sufficient hybridization of the enzymatic nucleic acid molecule to the target RNA and thus permit cleavage. One hundred percent complementarity is preferred, but complementarity as low as 50-75% may also be useful in this invention. The nucleic acids may be modified at the base, sugar, and/or phosphate groups. The term enzymatic nucleic acid is used interchangeably with phrases such as ribozymes, catalytic RNA, enzymatic RNA, catalytic DNA, aptazyme or aptamer-binding ribozyme, regulatable ribozyme, catalytic oligonucleotides, nucleozyme, DNAzyme, RNA enzyme, endoribonuclease, endonuclease, minizyme, leadzyme, oligozyme or DNA enzyme. All of these terminologies describe nucleic acid molecules with enzymatic activity. The specific enzymatic nucleic acid molecules described in the instant application are not meant to be limiting and those skilled in the art will recognize that all that is important in an enzymatic nucleic acid molecule of this invention is that it have a specific substrate binding site which is complementary to one or more of the target nucleic acid regions, and that it have nucleotide sequences within or surrounding that substrate binding site which impart a nucleic acid cleaving activity to the molecule (Cech et al., U.S. Pat. No. 4,987,071; Cech et al., 1988, JAMA).  
       [0031] By “nucleic acid molecule” as used herein is meant a molecule having nucleotides. The nucleic acid can be single, double, or multiple stranded and may comprise modified or unmodified nucleotides or non-nucleotides or various mixtures and combinations thereof.  
       [0032] By “enzymatic portion” or “catalytic domain” is meant that portion/region of the ribozyme essential for cleavage of a nucleic acid substrate (for example, see FIG. 1).  
       [0033] By “substrate binding arm” or “substrate binding domain” is meant that portion/region of a ribozyme which is complementary to (i.e., able to base-pair with) a portion of its substrate. Generally, such complementary is 100%, but can be less if desired. For example, as few as 10 bases out of 14 may be base-paired. Such arms are shown generally in FIGS. 1 and 3. That is, these arms contain sequences within a ribozyme which are intended to bring ribozyme and target RNA together through complementary base-pairing interactions. The ribozyme of the invention may have binding arms that are contiguous or non-contiguous and may be of varying lengths. The length of the binding arm(s) are preferably greater than or equal to four nucleotides; specifically 12-100 nucleotides; more specifically 14-24 nucleotides long. If two binding arms are chosen, the design is such that the length of the binding arms are symmetrical (i.e., each of the binding arms is of the same length; e.g., five and five nucleotides, six and six nucleotides or seven and seven nucleotides long) or asymmetrical (i.e., the binding arms are of different length; e.g., six and three nucleotides; three and six nucleotides long; four and five nucleotides long; four and six nucleotides long; four and seven nucleotides long; and the like).  
       [0034] By “Inozyme” motif is meant, an enzymatic nucleic acid molecule comprising a motif as described in Ludwig et al., U.S. Ser. No. 09/406,643, filed Sep. 27, 1999, entitled “COMPOSITIONS HAVING RNA CLEAVING ACTIVITY”, and International PCT publication Nos. WO 98/58058 and WO 98/58057, all incorporated by reference herein in their entirety including the drawings.  
       [0035] By “G-cleaver” motif is meant, an enzymatic nucleic acid molecule comprising a motif as described in Eckstein et al., International PCT publication No. WO 99/16871, incorporated by reference herein in its entirety including the drawings.  
       [0036] By “zinzyme” motif is meant, a class II enzymatic nucleic acid molecule comprising a motif as described in Beigelman et al., International PCT publication No. WO 99/55857, incorporated by reference herein in its entirety including the drawings. By “amberzyme” motif is meant, a class I enzymatic nucleic acid molecule comprising a motif as described in Beigelman et al., International PCT publication No. WO 99/55857, incorporated by reference herein in its entirety including the drawings.  
       [0037] By ‘DNAzyme’ is meant, an enzymatic nucleic acid molecule that does not require the presence of a 2′-OH group for its activity. In particular embodiments, the enzymatic nucleic acid molecule may have an attached linker(s) or other attached or associated groups, moieties, or chains containing one or more nucleotides with 2′-OH groups.  
       [0038] By “sufficient length” is meant an oligonucleotide of greater than or equal to 3 nucleotides that is of a length great enough to provide the intended function under the expected condition. For example, for binding arms of enzymatic nucleic acid “sufficient length” means that the binding arm sequence is long enough to provide stable binding to a target site under the expected binding conditions. Preferably, the binding arms are not so long as to prevent useful turnover.  
       [0039] By “stably interact” is meant, interaction of the oligonucleotides with target nucleic acid (e.g., by forming hydrogen bonds with complementary nucleotides in the target under physiological conditions).  
       [0040] By “equivalent” RNA to HCV is meant to include those naturally occurring RNA molecules associated with HCV infection in various animals, including human, rodent, primate, rabbit and pig. The equivalent RNA sequence also includes in addition to the coding region, regions such as 5′-untranslated region, 3′-untranslated region, introns, intron-exon junction and the like.  
       [0041] By “homology” is meant the nucleotide sequence of two or more nucleic acid molecules is partially or completely identical.  
       [0042] In one of the preferred embodiments of the inventions herein, the enzymatic nucleic acid molecule is formed in a hammerhead or hairpin motif, but may also be formed in the motif of a hepatitis d virus, group I intron, group II intron or RNaseP RNA (in association with an RNA guide sequence) or Neurospora VS RNA. Examples of such hammerhead motifs are described by Dreyfus, supra, Rossi et al., 1992,  AIDS Research and Human Retroviruses  8, 183; Hairpin motifs are described by Hampel et al., EP0360257, Hampel and Tritz, 1989  Biochemistry  28, 4929, Feldstein et al., 1989,  Gene  82, 53, Haseloff and Gerlach, 1989,  Gene,  82, 43, and Hampel et al., 1990  Nucleic Acids Res.  18, 299; The hepatitis d virus motif is described by Perrotta and Been, 1992  Biochemistry  31, 16; The RNaseP motif is described by Guerrier-Takada et al., 1983  Cell  35, 849; Forster and Altman, 1990,  Science  249, 783; Li and Altman, 1996,  Nucleic Acids Res.  24, 835; Neurospora VS RNA ribozyme motif is described by Collins (Saville and Collins, 1990  Cell  61, 685-696; Saville and Collins, 1991  Proc. Natl. Acad. Sci.  USA 88, 8826-8830; Collins and Olive, 1993  Biochemistry  32, 2795-2799; Guo and Collins, 1995,  EMBO. J.  14, 363); Group II introns are described by Griffin et al., 1995,  Chem. Biol.  2, 761; Michels and Pyle, 1995,  Biochemistry  34, 2965; Pyle et al., International PCT Publication No. WO 96/22689; The Group I intron is described by Cech et al., U.S. Pat. No. 4,987,071; and the DNAzyme motif is described by Chartrand et al., 1995,  Nucleic Acids Research  23, 4092; Santoro et al., 1997,  PNAS  94, 4262. These specific motifs are not limiting in the invention and those skilled in the art will recognize that all that is important in an enzymatic nucleic acid molecule of this invention is that it has a specific substrate binding site which is complementary to one or more of the target gene RNA regions, and that it have nucleotide sequences within or surrounding that substrate binding site which impart an RNA cleaving activity to the molecule.  
       [0043] By “complementarity” is meant that a nucleic acid can form hydrogen bond(s) with another RNA sequence by either traditional Watson-Crick or other non-traditional types. In reference to the nucleic molecules of the present invention, the binding free energy for a nucleic acid molecule with its target or complementary sequence is sufficient to allow the relevant function of the nucleic acid to proceed, e.g., ribozyme cleavage, antisense or triple helix inhibition. Determination of binding free energies for nucleic acid molecules is well known in the art (see, e.g., Turner et al., 1987,  CSH Symp. Quant. Biol.  LII pp.123-133; Frier et al., 1986,  Proc. Nat. Acad. Sci. USA  83:9373-9377; Turner et al., 1987,  J. Am. Chem. Soc.  109:3783-3785). A percent complementarity indicates the percentage of contiguous residues in a nucleic acid molecule which can form hydrogen bonds (e.g., Watson-Crick base pairing) with a second nucleic acid sequence (e.g., 5, 6, 7, 8, 9, 10 out of 10 being 50%, 60%, 70%, 80%, 90%, and 100% complementary). “Perfectly complementary” means that all the contiguous residues of a nucleic acid sequence will hydrogen bond with the same number of contiguous residues in a second nucleic acid sequence.  
       [0044] In a preferred embodiment, the invention provides a method for producing a class of enzymatic cleaving agents which exhibit a high degree of specificity for the RNA of a desired target. The enzymatic nucleic acid molecule is preferably targeted to a highly conserved sequence region of a target mRNAs encoding HCV proteins such that specific treatment of a disease or condition can be provided with either one or several enzymatic nucleic acids. Such enzymatic nucleic acid molecules can be delivered exogenously to specific cells as required. Alternatively, the ribozymes can be expressed from DNA/RNA vectors that are delivered to specific cells.  
       [0045] By “highly conserved sequence region” is meant, a nucleotide sequence of one or more regions in a target gene does not vary significantly from one generation to the other or from one biological system to the other.  
       [0046] Such ribozymes are useful for the prevention of the diseases and conditions discussed above, and any other diseases or conditions that are related to the levels of HCV activity in a cell or tissue.  
       [0047] By “related” is meant that the inhibition of HCV RNAs and thus reduction in the level respective viral activity will relieve to some extent the symptoms of the disease or condition.  
       [0048] In preferred embodiments, the ribozymes have binding arms which are complementary to the target sequences in Tables IV-VIII and X. Examples of such ribozymes are also shown in Tables IV-X. Examples of such ribozymes consist essentially of sequences defined in these tables. Other sequences may be present which do not interfere with such cleavage.  
       [0049] By “consists essentially of” is meant that the active ribozyme contains an enzymatic center or core equivalent to those in the examples, and binding arms able to bind mRNA such that cleavage at the target site occurs. Other sequences may be present which do not interfere with such cleavage. Thus, a core region may, for example, include one or more loop or stem-loop structures, which do not prevent enzymatic activity. “X” in the sequences in Tables V-VIII can be such a loop. A core sequence for a hammerhead ribozyme can be CUGAUGAG X CGAA where X=GCCGUUAGGC or other stem II region known in the art.  
       [0050] Thus, in a first aspect, the invention features ribozymes that inhibit gene expression and/or viral replication. These chemically or enzymatically synthesized RNA molecules contain substrate binding domains that bind to accessible regions of their target mRNAs. The RNA molecules also contain domains that catalyze the cleavage of RNA. The RNA molecules are preferably ribozymes of the hammerhead or hairpin motif. Upon binding, the ribozymes cleave the target mRNAs, preventing translation and protein accumulation. In the absence of the expression of the target gene, HCV gene expression and/or replication is inhibited.  
       [0051] In a preferred embodiment, ribozymes are added directly, or can be complexed with cationic lipids, packaged within liposomes, or otherwise delivered to target cells. The nucleic acid or nucleic acid complexes can be locally administered to relevant tissues ex vivo, or in vivo through injection, infusion pump or stent, with or without their incorporation in biopolymers. In another preferred embodiment, the ribozyme is administered to the site of HCV activity (e.g., hepatocytes) in an appropriate liposomal vehicle.  
       [0052] In another aspect of the invention, ribozymes that cleave target molecules and inhibit HCV activity are expressed from transcription units inserted into DNA or RNA vectors. The recombinant vectors are preferably DNA plasmids or viral vectors. Ribozyme expressing viral vectors could be constructed based on, but not limited to, adeno-associated virus, retrovirus, adenovirus, or alphavirus. Preferably, the recombinant vectors capable of expressing the ribozymes are delivered as described above, and persist in target cells. Alternatively, viral vectors may be used that provide for transient expression of ribozymes. Such vectors might be repeatedly administered as necessary. Once expressed, the ribozymes cleave the target mRNA. Delivery of ribozyme expressing vectors could be systemic, such as by intravenous or intramuscular administration, by administration to target cells ex-planted from the patient followed by reintroduction into the patient, or by any other means that would allow for introduction into the desired target cell (for a review see Couture and Stinchcomb, 1996,  TIG.,  12, 510). In another aspect of the invention, ribozymes that cleave target molecules and inhibit viral replication are expressed from transcription units inserted into DNA, RNA, or viral vectors. Preferably, the recombinant vectors capable of expressing the ribozymes are locally delivered as described above, and transiently persist in smooth muscle cells. However, other mammalian cell vectors that direct the expression of RNA may be used for this purpose.  
       [0053] By “patient” is meant an organism which is a donor or recipient of explanted cells or the cells themselves. “Patient” also refers to an organism to which enzymatic nucleic acid molecules can be administered. Preferably, a patient is a mammal or mammalian cells. More preferably, a patient is a human or human cells.  
       [0054] As used herein “cell” is used in its usual biological sense, and does not refer to an entire multicellular organism, e.g., specifically does not refer to a human. The cell may be present in an organism which may be a human but is preferably a non-human multicellular organism, e.g., birds, plants and mammals such as cows, sheep, apes, monkeys, swine, dogs, and cats. The cell may be prokaryotic (e.g. bacterial cell) or eukaryotic (e.g., mammalian or plant cell).  
       [0055] By RNA is meant a molecule comprising at least one ribonucleotide residue. By “ribonucleotide” is meant a nucleotide with a hydroxyl group at the 2′ position (eg; 2′-OH) of a β-D-ribo-furanose moiety.  
       [0056] By “vectors” is meant any nucleic acid- and/or viral-based technique used to deliver a desired nucleic acid.  
       [0057] These ribozymes, individually, or in combination or in conjunction with other drugs, can be used to treat diseases or conditions discussed above. For example, to treat a disease or condition associated with HCV levels, the patient may be treated, or other appropriate cells may be treated, as is evident to those skilled in the art.  
       [0058] In a further embodiment, the described molecules can be used in combination with other known treatments to treat conditions or diseases discussed above. For example, the described molecules could be used in combination with one or more known therapeutic agents to treat liver failure, hepatocellular carcinoma, cirrhosis, and/or other disease states associated with HCV infection. Additional known therapeutic agents are those comprising antivirals, interferon, and/or antisense compounds.  
       [0059] By “comprising” is meant including, but not limited to, whatever follows the word “comprising”. Thus, use of the term “comprising” indicates that the listed elements are required or mandatory, but that other elements are optional and may or may not be present. By “consisting of” is meant including, and limited to, whatever follows the phrase “consisting of”. Thus, the phrase “consisting of” indicates that the listed elements are required or mandatory, and that no other elements may be present. By “consisting essentially of” is meant including any elements listed after the phrase, and limited to other elements that do not interfere with or contribute to the activity or action specified in the disclosure for the listed elements. Thus, the phrase “consisting essentially of” indicates that the listed elements are required or mandatory, but that other elements are optional and may or may not be present depending upon whether or not they affect the activity or action of the listed elements.  
       [0060] Other features and advantages of the invention will be apparent from the following description of the preferred embodiments thereof, and from the claims. 
     
    
    
     DESCRIPTION OF THE PREFERRED EMBODIMENTS  
     [0061] The drawings will first briefly be described.  
     [0062] Drawings  
     [0063]FIG. 1 shows the secondary structure model for seven different classes of enzymatic nucleic acid molecules. Arrow indicates the site of cleavage. --------- indicate the target sequence. Lines interspersed with dots are meant to indicate tertiary interactions. —is meant to indicate base-paired interaction. Group I Intron: P1-P9.0 represent various stem-loop structures (Cech et al., 1994,  Nature Struc. Bio.,  1, 273).  
     [0064] RNase P (M 1 RNA): EGS represents external guide sequence (Forster et al., 1990,  Science,  249, 783; Pace et al., 1990,  J. Biol. Chem.,  265, 3587). Group II Intron: 5′SS means 5′ splice site; 3′SS means 3′-splice site; IBS means intron binding site; EBS means exon binding site (Pyle et al., 1994,  Biochemistry,  33, 2716). VS RNA: I-VI are meant to indicate six stem-loop structures; shaded regions are meant to indicate tertiary interaction (Collins, International PCT Publication No. WO 96/19577). HDV Ribozyme: : I-IV are meant to indicate four stem-loop structures (Been et al., U.S. Pat. No. 5,625,047). Hammerhead Ribozyme: : I-III are meant to indicate three stem-loop structures; stems I-III can be of any length and may be symmetrical or asymmetrical (Usman et al., 1996,  Curr. Op. Struct. Bio.,  1, 527). Hairpin Ribozyme: Helix 1, 4 and 5 can be of any length; Helix 2 is between 3 and 8 base-pairs long; Y is a pyrimidine; Helix 2 (H2) is provided with a least 4 base pairs (i.e., n is 1, 2, 3 or 4) and helix 5 can be optionally provided of length 2 or more bases (preferably 3-20 bases, i.e., m is from 1-20 or more). Helix 2 and helix 5 may be covalently linked by one or more bases (i.e., r is 1 base). Helix 1, 4 or 5 may also be extended by 2 or more base pairs (e.g., 4-20 base pairs) to stabilize the ribozyme structure, and preferably is a protein binding site. In each instance, each N and N′ independently is any normal or modified base and each dash represents a potential base-pairing interaction. These nucleotides may be modified at the sugar, base or phosphate. Complete base-pairing is not required in the helices, but is preferred. Helix 1 and 4 can be of any size (i.e., o and p is each independently from 0 to any number, e.g., 20) as long as some base-pairing is maintained. Essential bases are shown as specific bases in the structure, but those in the art will recognize that one or more may be modified chemically (abasic, base, sugar and/or phosphate modifications) or replaced with another base without significant effect. Helix 4 can be formed from two separate molecules, i.e., without a connecting loop. The connecting loop when present may be a ribonucleotide with or without modifications to its base, sugar or phosphate. “q” is ≧2 bases. The connecting loop can also be replaced with a non-nucleotide linker molecule. H refers to bases A, U, or C. Y refers to pyrimidine bases. “_” refers to a covalent bond. (Burke et al., 1996,  Nucleic Acids &amp; Mol. Biol.,  10, 129; Chowrira et al., U.S. Pat. No. 5,631,359).  
     [0065]FIG. 2 is a graph displaying the ability of ribozymes targeting various sites within the conserved 5′HCV UTR region to cleave the transcripts made from several genotypes.  
     [0066]FIG. 3 is a schematic representation of the Dual Reporter System utilized to demonstrate ribozyme-mediated reduction of luciferase activity in cell culture.  
     [0067]FIG. 4 is a graph demonstrating the ability of ribozymes to reduce luciferase activity in OST-7 cells.  
     [0068]FIG. 5 is a graph demonstrating the ability of ribozymes targeting sites HCV 0.5-313 and HCV 0.5-318, to reduce luciferase activity in OST-7 cells compared to their inactive controls.  
     [0069]FIG. 6A is a bar graph demonstrating the effect of ribozyme treatment on HCV-Polio virus (PV) replication. HeLa cells in 96-well plates were infected with HCV-PV at a multiplicity of infection (MOI) of 0.1. Virus inoculum was then replaced with media containing 5% serum and ribozyme or control (200nM), as indicated, complexed to a cationic lipid. After 24 hours, cells were lysed 3 times by freeze/thaw and virus was quantified by plaque assay. Scrambled control (SAC), binding control (BAC), 3 P=S ribozymes, and 4 P=S ribozymes are indicated. Plaque forming units (pfu)/ml are shown as the mean of triplicate samples +standard deviation (S.D.).  
     [0070]FIG. 6B is a bar graph demonstrating the effect of ribozyme treatment on wild type PV replication. HeLa cells in 96-well plates were infected with wild type PV at an MOI=0.05 for 30 minutes. All ribozymes contained 4P=S in (B). Plaque forming units (pfu)/ml are shown as the mean of triplicate samples+standard deviation (S.D.).  
     [0071]FIG. 7 is a schematic representation of various hammerhead ribozyme constructs targeted against HCV RNA.  
     [0072]FIG. 8 is a graph demonstrating the effect of site 183 ribozyme treatment on a single round of HCV-PV infection. HeLa cells were infected with HCV-PV at an MOI=5 for 30 minutes prior to treatment with ribozymes or control. Cells were lysed after 6, 7, or 8 hours and virus was quantified by plaque assay. Ribozyme binding arm/stem II formats (7/4, 7/3, 6/4, 6/3) and scrambled control (SAC, 7/4 format) are indicated. All contained 4P=S stabilization. Results in pfu/ml are shown as the median of duplicate samples±range.  
     [0073]FIG. 9 shows the secondary structure models of three ribozyme motifs described in this application.  
     [0074]FIG. 10 shows the activity of anti-HCV ribozymes in combination with Interferon. Results in pfu/ml are shown as the median of duplicate samples±range. BAC, binding attenuated control molecule; IF, interferon; Rz, hammerhead ribozyme targeted to HCV site 183; pfu, plaque forming unit.  
     [0075]FIG. 11 is a bar graph demonstrating the effect of ribozyme treatment on HCV-Polio virus (PV) replication using anti-HCV ribozymes directed against sites in the HCV minus strand. Both RPI motif I (Hammerhead) and motif II (Inozyme) ribozymes are represented. HeLa cells in 96-well plates were infected with HCV-PV at a multiplicity of infection (MOI) of 0.1. Virus inoculum was then replaced with media containing 5% serum and ribozyme or control (200 nM), as indicated, complexed to a cationic lipid. After 24 hours, cells were lysed 3 times by freeze/thaw and virus was quantified by plaque assay. Scrambled control (SAC) and ribozymes targeting different sites are indicated. Plaque forming units (pfu)/ml are shown as the mean of triplicate samples+standard deviation (S.D.). Ribozymes used in this study are shown in Table X.  
     [0076]FIG. 12 is a bar graph demonstrating the effect of ribozyme treatment on HCV-Polio virus (PV) replication using anti-HCV ribozymes directed against additional sites in the HCV minus strand. Both RPI motif I and motif II ribozymes are represented. HeLa cells in 96-well plates were infected with HCV-PV at a multiplicity of infection (MOI) of 0.1. Virus inoculum was then replaced with media containing 5% serum and ribozyme or control (200 nM), as indicated, complexed to a cationic lipid. After 24 hours cells, were lysed 3 times by freeze/thaw and virus was quantified by plaque assay. Scrambled control (SAC) and ribozymes targeting different sites are indicated. Plaque forming units (pfu)/ml are shown as the mean of triplicate samples+standard deviation (S.D.). Ribozymes used in this study are shown in Table X.  
     [0077]FIG. 13 is a bar graph showing the dose response of a HCV minus strand site 205 directed anti-HCV ribozyme (RPI No. 15006, Table X). Plaque forming units (pfu)/ml are shown as the mean of triplicate samples+standard deviation (S.D.). Results are shown in plaque forming units (pfu)/ml vs. ribozyme concentration in nM.  
     [0078]FIG. 14 is a graph showing the dose response of a HCV plus strand site 195 directed anti-HCV ribozyme (RPI No. 13919) when mixed with differing anti-HCV minus strand directed ribozymes (Table X). Results are shown in plaque forming units (pfu)/ml vs. ribozyme concentration in nM. 
    
    
     [0079] Ribozymes  
     [0080] Seven basic varieties of naturally-occurring enzymatic RNAs are known presently. In addition, several in vitro selection (evolution) strategies (Orgel, 1979,  Proc. R. Soc. London,  B 205, 435) have been used to evolve new nucleic acid catalysts capable of catalyzing cleavage and ligation of phosphodiester linkages (Joyce, 1989,  Gene,  82, 83-87; Beaudry et al., 1992,  Science  257, 635-641; Joyce, 1992,  Scientific American  267, 90-97; Breaker et al., 1994,  TIBTECH  12, 268; Bartel et al.,1993,  Science  261:1411-1418; Szostak, 1993,  TIBS  17, 89-93; Kumar et al., 1995,  FASEB J.,  9, 1183; Breaker, 1996,  Curr. Op. Biotech.,  7, 442; Santoro et al., 1997,  Proc. Natl. Acad. Sci.,  94, 4262; Tang et al., 1997,  RNA  3, 914; Nakamaye &amp; Eckstein, 1994, supra; Long &amp; Uhlenbeck, 1994, supra; Ishizaka et al., 1995, supra; Vaish et al., 1997,  Biochemistry  36, 6495; all of these publications are incorporated by reference herein). Each can catalyze a series of reactions including the hydrolysis of phosphodiester bonds in trans (and thus can cleave other RNA molecules) under physiological conditions. Table I summarizes some of the characteristics of some of these ribozymes. In general, enzymatic nucleic acids act by first binding to a target RNA. Such binding occurs through the target binding portion of an enzymatic nucleic acid which is held in close proximity to an enzymatic portion of the molecule that acts to cleave the target RNA. Thus, the enzymatic nucleic acid first recognizes and then binds a target RNA through complementary base-pairing, and once bound to the correct site, acts enzymatically to cut the target RNA. Strategic cleavage of such a target RNA will destroy its ability to direct synthesis of an encoded protein. After an enzymatic nucleic acid has bound and cleaved its RNA target, it is released from that RNA to search for another target and can repeatedly bind and cleave new targets.  
     [0081] The enzymatic nature of a ribozyme is advantageous over other technologies, since the concentration of ribozyme necessary to affect a therapeutic treatment is lower. This advantage reflects the ability of the ribozyme to act enzymatically. Thus, a single ribozyme molecule is able to cleave many molecules of target RNA. In addition, the ribozyme is a highly specific inhibitor, with the specificity of inhibition depending not only on the base-pairing mechanism of binding to the target RNA, but also on the mechanism of target RNA cleavage. Single mismatches, or base-substitutions, near the site of cleavage can be chosen to completely eliminate catalytic activity of a ribozyme.  
     [0082] Nucleic acid molecules having an endonuclease enzymatic activity are able to repeatedly cleave other separate RNA molecules in a nucleotide base sequence-specific manner. Such enzymatic nucleic acid molecules can be targeted to virtually any RNA transcript, and efficient cleavage achieved in vitro (Zaug et al., 324,  Nature  429 1986; Uhlenbeck, 1987  Nature  328, 596; Kim et al., 84  Proc. Natl. Acad. Sci. USA  8788, 1987; Dreyfus, 1988,  Einstein Quart. J. Bio. Med.,  6, 92; Haseloff and Gerlach, 334  Nature  585, 1988; Cech, 260  JAMA  3030, 1988; and Jefferies et al., 17  Nucleic Acids Research  1371, 1989; Chartrand et al., 1995,  Nucleic Acids Research  23, 4092; Santoro et al., 1997,  PNAS  94, 4262).  
     [0083] Because of their sequence-specificity, trans-cleaving ribozymes show promise as therapeutic agents for human disease (Usman &amp; McSwiggen, 1995  Ann. Rep. Med. Chem.  30, 285-294; Christoffersen and Marr, 1995  J. Med. Chem.  38, 2023-2037). Ribozymes can be designed to cleave specific RNA targets within the background of cellular RNA. Such a cleavage event renders the RNA non-functional and abrogates protein expression from that RNA. In this manner, synthesis of a protein associated with a disease state can be selectively inhibited.  
     [0084] Ribozymes that cleave the specified sites in HCV RNAs represent a novel therapeutic approach to infection by the hepatitis C virus. Applicant indicates that ribozymes are able to inhibit the activity of HCV and that the catalytic activity of the ribozymes is required for their inhibitory effect. Those of ordinary skill in the art will find that it is clear from the examples described that other ribozymes that cleave HCV RNAs may be readily designed and are within the invention.  
     [0085] Target sites  
     [0086] Targets for useful ribozymes can be determined as disclosed in Draper et al., WO 93/23569; Sullivan et al., WO 93/23057; Thompson et al., WO 94/02595; Draper et al., WO 95/04818; McSwiggen et al., U.S. Pat. No. 5,525,468; and, are all hereby incorporated by reference herein in their totalities. Rather than repeat the guidance provided in those documents here, below are provided specific examples of such methods, not limiting to those in the art. Ribozymes to such targets are designed as described in those applications and synthesized to be tested in vitro and in vivo, as also described. Such ribozymes can also be optimized and delivered as described therein.  
     [0087] The sequence of HCV RNAs were screened for optimal ribozyme target sites using a computer folding algorithm. Hammerhead or hairpin ribozyme cleavage sites were identified. These sites are shown in Tables IV-VIII and X (All sequences are 5′ to 3′ in the tables). The nucleotide base position is noted in the tables as that site to be cleaved by the designated type of ribozyme. The nucleotide base position is noted in the tables as that site to be cleaved by the designated type of ribozyme.  
     [0088] Because HCV RNAs are highly homologous in certain regions, some ribozyme target sites are also homologous (see Table IV and VIII). In this case, a single ribozyme will target different classes of HCV RNA. The advantage of one ribozyme that targets several classes of HCV RNA is clear, especially in cases where one or more of these RNAs may contribute to the disease state.  
     [0089] Hammerhead or hairpin ribozymes were designed that could bind and were individually analyzed by computer folding (Jaeger et al., 1989  Proc. Natl. Acad. Sci. USA,  86, 7706) to assess whether the ribozyme sequences fold into the appropriate secondary structure. Those ribozymes with unfavorable intramolecular interactions between the binding arms and the catalytic core are eliminated from consideration. Varying binding arm lengths can be chosen to optimize activity. Generally, at least 5 bases on each arm are able to bind to, or otherwise interact with, the target RNA. Ribozymes of the hammerhead or hairpin motif were designed to anneal to various sites in the mRNA message. The binding arms are complementary to the target site sequences described above.  
     [0090] Ribozyme Synthesis  
     [0091] Synthesis of nucleic acids greater than 100 nucleotides in length is difficult using automated methods, and the therapeutic cost of such molecules is prohibitive. In this invention, small nucleic acid motifs (“small refers to nucleic acid motifs no more than 100 nucleotides in length, preferably no more than 80 nucleotides in length, and most preferably no more than 50 nucleotides in length; e.g., antisense oligonucleotides, hammerhead or the Inozyme ribozymes) are preferably used for exogenous delivery. The simple structure of these molecules increases the ability of the nucleic acid to invade targeted regions of RNA structure. Exemplary molecules of the instant invention are chemically synthesized, and others can similarly be synthesized.  
     [0092] The method of synthesis used for normal RNA including certain enzymatic nucleic acid molecules follows the procedure as described in Usman et al., 1987,  J. Am. Chem. Soc.,  109, 7845; Scaringe et al., 1990,  Nucleic Acids Res.,  18, 5433; Wincott et al., 1995,  Nucleic Acids Res.  23, 2677-2684; and Wincott et al., 1997,  Methods Mol. Bio.,  74, 59, and makes use of common nucleic acid protecting and coupling groups, such as dimethoxytrityl at the 5′-end, and phosphoramidites at the 3′-end. In a non-limiting example, small scale syntheses are conducted on a 394 Applied Biosystems, Inc. synthesizer using a 0.2 μmol scale protocol with a 7.5 min coupling step for alkylsilyl protected nucleotides and a 2.5 min coupling step for 2′-O-methylated nucleotides. Table II outlines the amounts and the contact times of the reagents used in the synthesis cycle. Alternatively, syntheses at the 0.2 μmol scale can be done on a 96-well plate synthesizer, such as the instrument produced by Protogene (Palo Alto, Calif.) with minimal modification to the cycle. A 33-fold excess (60 μL of 0.11 M=6.6 μmol) of 2′-O-methyl phosphoramidite and a 75-fold excess of S-ethyl tetrazole (60 μL of 0.25 M=15 μmol) can be used in each coupling cycle of 2′-O-methyl residues relative to polymer-bound 5′-hydroxyl. A 66-fold excess (120 μL of 0.11 M=13.2 μmol) of alkylsilyl (ribo) protected phosphoramidite and a 150-fold excess of S-ethyl tetrazole (120 μL of 0.25 M=30 μmol) can be used in each coupling cycle of ribo residues relative to polymer-bound 5′-hydroxyl. Average coupling yields on the 394 Applied Biosystems, Inc. synthesizer, determined by colorimetric quantitation of the trityl fractions, are typically 97.5-99%. Other oligonucleotide synthesis reagents for the 394 Applied Biosystems, Inc. synthesizer include; detritylation solution is 3% TCA in methylene chloride (ABI); capping is performed with 16% N-methyl imidazole in THF (ABI) and 10% acetic anhydride/10% 2,6-lutidine in THF (ABI); oxidation solution is 16.9 mM I 2 , 49 mM pyridine, 9% water in THF (PERSEPTIVE™). Burdick &amp; Jackson Synthesis Grade acetonitrile is used directly from the reagent bottle. S-Ethyltetrazole solution (0.25 M in acetonitrile) is made up from the solid obtained from American International Chemical, Inc. Alternately, for the introduction of phosphorothioate linkages, Beaucage reagent (3H-1,2-Benzodithiol-3-one 1,1-dioxide0.05 M in acetonitrile) is used.  
     [0093] Deprotection of the RNA is performed using either a two-pot or one-pot protocol. For the two-pot protocol, the polymer-bound trityl-on oligoribonucleotide is transferred to a 4 mL glass screw top vial and suspended in a solution of 40% aq. methylamine (1 mL) at 65° C. for 10 min. After cooling to −20° C. the supernatant is removed from the polymer support. The support is washed three times with 1.0 mL of EtOH:MeCN:H2O/3:1:1, vortexed and the supernatant is then added to the first supernatant. The combined supernatants, containing the oligoribonucleotide, are dried to a white powder. The base deprotected oligoribonucleotide is resuspended in anhydrous TEA/HF/NMP solution (300 μL of a solution of 1.5 mL N-methylpyrrolidinone, 750 μL TEA and 1 mL TEA·3HF to provide a 1.4 M HF concentration) and heated to 65° C. After 1.5 h, the oligomer is quenched with 1.5 M NH 4 HCO 3 .  
     [0094] Alternatively, for the one-pot protocol, the polymer-bound trityl-on oligoribonucleotide is transferred to a 4 mL glass screw top vial and suspended in a solution of 33% ethanolic methylamine/DMSO: 1/1 (0.8 mL) at 65° C. for 15 min. The vial is brought to r.t. TEA·3HF (0.1 mL) is added and the vial is heated at 65° C. for 15 min. The sample is cooled at −20° C. and then quenched with 1.5 M NH 4 HCO 3 .  
     [0095] For anion exchange desalting of the deprotected oligomer, the TEAB solution was loaded onto a Qiagen 500® anion exchange cartridge (Qiagen Inc.) that was prewashed with 50 mM TEAB (10 mL). After washing the loaded cartridge with 50 mM TEAB (10 mL), the RNA was eluted with 2 M TEAB (10 mL) and dried down to a white powder.  
     [0096] For purification of the trityl-on oligomers, the quenched NH 4 HCO 3  solution is loaded onto a C- 18  containing cartridge that had been prewashed with acetonitrile followed by 50 mM TEAA. After washing the loaded cartridge with water, the RNA is detritylated with 0.5% TFA for 13 min. The cartridge is then washed again with water, salt exchanged with 1 M NaCl and washed with water again. The oligonucleotide is then eluted with 30% acetonitrile.  
     [0097] Inactive hammerhead ribozymes were synthesized by substituting switching the order of G 5 A 6  and substituting a U for A, 14  (numbering from Hertel, K. J., et al., 1992,  Nucleic Acids Res.,  20, 3252). Inactive ribozymes were also by synthesized by substituting a U for G 5  and a U for A 14 . In some cases, the sequence of the substrate binding arms were randomized while the overall base composition was maintained.  
     [0098] The average stepwise coupling yields are typically &gt;98% (Wincott et al., 1995  Nucleic Acids Res.  23, 2677-2684). Those of ordinary skill in the art will recognize that the scale of synthesis can be adapted to be larger or smaller than the example described above including but not limited to 96-well format, all that is important is the ratio of chemicals used in the reaction.  
     [0099] Hairpin ribozymes are synthesized in two parts and annealed to reconstruct the active ribozyme (Chowrira and Burke, 1992  Nucleic Acids Res.,  20, 2835-2840). Ribozymes are also synthesized from DNA templates using bacteriophage T7 RNA polymerase (Milligan and Uhlenbeck, 1989,  Methods Enzymol.  180, 51).  
     [0100] Alternatively, the nucleic acid molecules of the present invention can be synthesized separately and joined together post-synthetically, for example by ligation (Moore et al., 1992,  Science  256, 9923; Draper et al., International PCT publication No. WO 93/23569; Shabarova et al., 1991,  Nucleic Acids Research  19, 4247; Bellon et al., 1997,  Nucleosides &amp; Nucleotides,  16, 951;  Bellon et al.,  1997, Bioconjugate Chem. 8, 204).  
     [0101] The nucleic acid molecules of the present invention are modified extensively to enhance stability by modification with nuclease resistant groups, for example, 2′-amino, 2′-C-allyl, 2′-flouro, 2′-O-methyl, 2′-H (for a review see Usman and Cedergren, 1992, 30  TIBS  17, 34; Usman et al., 1994,  Nucleic Acids Symp. Ser.  31, 163). Ribozymes are purified by gel electrophoresis using general methods or are purified by high pressure liquid chromatography (HPLC; see Wincott et al., supra, the totality of which is hereby incorporated herein by reference) and are re-suspended in water.  
     [0102] The sequences of the ribozymes that are chemically synthesized, useful in this study, are shown in Tables IV to X. Those in the art will recognize that these sequences are representative only of many more such sequences where the enzymatic portion of the ribozyme (all but the binding arms) is altered to affect activity. The ribozyme sequences listed in Tables IV to X may be formed of ribonucleotides or other nucleotides or non-nucleotides. Such ribozymes with enzymatic activity are equivalent to the ribozymes described specifically in the tables.  
     [0103] Optimizing Activity of the nucleic acid molecule of the invention.  
     [0104] Chemically synthesizing nucleic acid molecules with modifications (base, sugar and/or phosphate) that prevent their degradation by serum ribonucleases may increase their potency (see e.g., Eckstein et al., International Publication No. WO 92/07065; Perrault et al., 1990  Nature  344, 565; Pieken et al., 1991,  Science  253, 314; Usman and Cedergren, 1992,  Trends in Biochem. Sci.  17, 334; Usman et al., International Publication No. WO 93/15187; and Rossi et al., International Publication No. WO 91/03162; Sproat, U.S. Pat. No. 5,334,711; and Burgin et al., supra; all of these describe various chemical modifications that can be made to the base, phosphate and/or sugar moieties of the nucleic acid molecules herein). All these publications are hereby incorporated by reference herein. Modifications which enhance their efficacy in cells, and removal of bases from nucleic acid molecules to shorten oligonucleotide synthesis times and reduce chemical requirements are desired.  
     [0105] There are several examples in the art describing sugar, base and phosphate modifications that can be introduced into nucleic acid molecules with significant enhancement in their nuclease stability and efficacy. For example, oligonucleotides are modified to enhance stability and/or enhance biological activity by modification with nuclease resistant groups, for example, 2′-amino, 2′-C-allyl, 2′-flouro, 2′-O-methyl, 2′-H, nucleotide base modifications (for a review, see Usman and Cedergren, 1992,  TIBS.  17, 34; Usman et al., 1994,  Nucleic Acids Symp. Ser.  31, 163; Burgin et al., 1996,  Biochemistry,  35, 14090). All of these publications are incorporated by reference herein. Sugar modification of nucleic acid molecules have been extensively described in the art (see Eckstein et al., International Publication PCT No. WO 92/07065; Perrault et al.  Nature,  1990, 344, 565-568; Pieken et al.  Science,  1991, 253, 314-317; Usman and Cedergren,  Trends in Biochem. Sci.,  1992, 17, 334-339; Usman et al. International Publication PCT No. WO 93/15187; Sproat, U.S. Pat. No. 5,334,711 and Beigelman et al., 1995,  J. Biol. Chem.,  270, 25702; Beigelman et al., International PCT publication No. WO 97/26270; Beigelman et al., U.S. Pat. No. 5,716,824; Usman et al, U.S. Pat. No. 5,627,053; Woolf et al., International PCT Publication No. WO 98/13526; Thompson et al., U.S. Ser. No. 60/082,404 which was filed on Apr. 20, 1998; Karpeisky et al., 1998,  Tetrahedron Lett.,  39, 1131; Earnshaw and Gait, 1998,  Biopolymers  ( Nucleic acid Sciences ), 48, 39-55; Verma and Eckstein, 1998,  Annu. Rev. Biochem.,  67, 99-134; and Burlina et al., 1997,  Bioorg. Med. Chem.,  5, 1999-2010; all of the references are hereby incorporated in their totality by reference herein). Such publications describe general methods and strategies to determine the location of incorporation of sugar, base and/or phosphate modifications and the like into ribozymes without inhibiting catalysis, and are incorporated by reference herein. In view of such teachings, similar modifications can be used as described herein to modify the nucleic acid molecules of the instant invention.  
     [0106] While chemical modification of oligonucleotide internucleotide linkages with phosphorothioate, phosphorothioate, and/or 5′-methylphosphonate linkages improves stability, too many of these modifications may cause some toxicity. Therefore when designing nucleic acid molecules the amount of these internucleotide linkages should be minimized. The reduction in the concentration of these linkages should lower toxicity resulting in increased efficacy and higher specificity of these molecules.  
     [0107] Nucleic acid molecules having chemical modifications which maintain or enhance activity are provided. Such nucleic acid is also generally more resistant to nucleases than unmodified nucleic acid. Thus, in a cell and/or in vivo the activity may not be significantly lowered. Therapeutic nucleic acid molecules delivered exogenously must optimally be stable within cells until translation of the target RNA has been inhibited long enough to reduce the levels of the undesirable protein. This period of time varies between hours to days depending upon the disease state. Clearly, nucleic acid molecules must be resistant to nucleases in order to function as effective intracellular therapeutic agents. Improvements in the chemical synthesis of RNA and DNA (Wincott et aL, 1995  Nucleic Acids Res.  23, 2677; Caruthers et al., 1992,  Methods in Enzymology  211,3-19 (incorporated by reference herein) have expanded the ability to modify nucleic acid molecules by introducing nucleotide modifications to enhance their nuclease stability as described above.  
     [0108] Use of these the nucleic acid-based molecules of the invention will lead to better treatment of the disease progression by affording the possibility of combination therapies (e.g., multiple antisense or enzymatic nucleic acid molecules targeted to different genes, nucleic acid molecules coupled with known small molecule inhibitors, or intermittent treatment with combinations of molecules (including different motifs) and/or other chemical or biological molecules). The treatment of patients with nucleic acid molecules may also include combinations of different types of nucleic acid molecules.  
     [0109] Therapeutic nucleic acid molecules (e.g., enzymatic nucleic acid molecules) delivered exogenously must optimally be stable within cells until translation of the target RNA has been inhibited long enough to reduce the levels of the undesirable protein. This period of time varies between hours to days depending upon the disease state. Clearly, these nucleic acid molecules must be resistant to nucleases in order to function as effective intracellular therapeutic agents. Improvements in the chemical synthesis of nucleic acid molecules described in the instant invention and in the art have expanded the ability to modify nucleic acid molecules by introducing nucleotide modifications to enhance their nuclease stability as described above.  
     [0110] By “enhanced enzymatic activity” is meant to include activity measured in cells and/or in vivo where the activity is a reflection of both catalytic activity and ribozyme stability. In this invention, the product of these properties is increased or not significantly (less that 10 fold) decreased in vivo compared to an all RNA ribozyme or all DNA enzyme.  
     [0111] In yet another preferred embodiment, nucleic acid catalysts having chemical modifications which maintain or enhance enzymatic activity is provided. Such nucleic acid is also generally more resistant to nucleases than unmodified nucleic acid. Thus, in a cell and/or in vivo the activity may not be significantly lowered. As exemplified herein such ribozymes are useful in a cell and/or in vivo even if activity over all is reduced 10-fold (Burgin et al., 1996,  Biochemistry,  35, 14090). Such ribozymes herein are said to “maintain” the enzymatic activity of an all RNA ribozyme.  
     [0112] In another aspect the nucleic acid molecules comprise a 5′and/or a 3′-cap structure.  
     [0113] By “cap structure” is meant chemical modifications, which have been incorporated at either terminus of the oligonucleotide (see, for example, Wincott et al., WO 97/26270, incorporated by reference herein). These terminal modifications protect the nucleic acid molecule from exonuclease degradation, and may help in delivery and/or localization within a cell. The cap may be present at the 5′-terminus (5′-cap) or at the 3′-terminus (3′-cap) or may be present on both termini. In non-limiting examples: the 5′-cap is selected from the group comprising inverted abasic residue (moiety), 4′,5′-methylene nucleotide; 1-(beta-D-erythrofuranosyl) nucleotide, 4′-thio nucleotide, carbocyclic nucleotide; 1,5-anhydrohexitol nucleotide; L-nucleotides; alpha-nucleotides; modified base nucleotide; phosphorodithioate linkage; threo-pentofuranosyl nucleotide; acyclic 3′,4′-seco nucleotide; acyclic 3,4-dihydroxybutyl nucleotide; acyclic 3,5-dihydroxypentyl nucleotide, 3′-3′-inverted nucleotide moiety; 3′-3′-inverted abasic moiety; 3′-2′-inverted nucleotide moiety; 3′-2′-inverted abasic moiety; 1,4-butanediol phosphate; 3′-phosphoramidate; hexylphosphate; aminohexyl phosphate; 3′-phosphate; 3′-phosphorothioate; phosphorodithioate; or bridging or non-bridging methylphosphonate moiety (for more details see Wincott et al., International PCT publication No. WO 97/26270, incorporated by reference herein).  
     [0114] In yet another preferred embodiment, the 3′-cap is selected from a group comprising, 4′,5′-methylene nucleotide; 1-(beta-D-erythrofuranosyl) nucleotide; 4′-thio nucleotide, carbocyclic nucleotide; 5′-amino-alkyl phosphate; 1,3-diamino-2-propyl phosphate, 3-aminopropyl phosphate; 6-aminohexyl phosphate; 1,2-aminododecyl phosphate; hydroxypropyl phosphate; 1,5-anhydrohexitol nucleotide; L-nucleotide; alpha-nucleotide; modified base nucleotide; phosphorodithioate; threo-pentofuranosyl nucleotide; acyclic 3′,4′-seco nucleotide; 3,4-dihydroxybutyl nucleotide; 3,5-dihydroxypentyl nucleotide, 5′-5′-inverted nucleotide moiety; 5′-5′-inverted abasic moiety; 5′-phosphoramidate; 5′-phosphorothioate; 1,4-butanediol phosphate; 5′-amino; bridging and/or non-bridging 5′-phosphoramidate, phosphorothioate and/or phosphorodithioate, bridging or non bridging methylphosphonate and 5′-mercapto moieties (for more details, see Beaucage and Iyer, 1993,  Tetrahedron  49, 1925; incorporated by reference herein). By the term “non-nucleotide” is meant any group or compound which can be incorporated into a nucleic acid chain in the place of one or more nucleotide units, including either sugar and/or phosphate substitutions, and allows the remaining bases to exhibit their enzymatic activity. The group or compound is abasic in that it does not contain a commonly recognized nucleotide base, such as adenosine, guanine, cytosine, uracil or thymine.  
     [0115] An “alkyl” group refers to a saturated aliphatic hydrocarbon, including straight-chain, branched-chain, and cyclic alkyl groups. Preferably, the alkyl group has 1 to 12 carbons. More preferably it is a lower alkyl of from 1 to 7 carbons, more preferably 1 to 4 carbons. The alkyl group may be substituted or unsubstituted. When substituted the substituted group(s) is preferably, hydroxyl, cyano, alkoxy, ═O, ═S, NO 2  or N(CH 3 ) 2,  amino, or SH. The term also includes alkenyl groups which are unsaturated hydrocarbon groups containing at least one carbon-carbon double bond, including straight-chain, branched-chain, and cyclic groups. Preferably, the alkenyl group has 1 to 12 carbons. More preferably it is a lower alkenyl of from 1 to 7 carbons, more preferably 1 to 4 carbons. The alkenyl group may be substituted or unsubstituted. When substituted the substituted group(s) is preferably, hydroxyl, cyano, alkoxy, ═O, ═S, NO 2,  halogen, N(CH 3 ) 2,  amino, or SH. The term “alkyl” also includes alkynyl groups which have an unsaturated hydrocarbon group containing at least one carbon-carbon triple bond, including straight-chain, branched-chain, and cyclic groups. Preferably, the alkynyl group has 1 to 12 carbons. More preferably it is a lower alkynyl of from 1 to 7 carbons, more preferably 1 to 4 carbons. The alkynyl group may be substituted or unsubstituted. When substituted the substituted group(s) is preferably, hydroxyl, cyano, alkoxy, ═O, ═S, NO 2  or N(CH 3 ) 2 , amino or SH.  
     [0116] Such alkyl groups may also include aryl, alkylaryl, carbocyclic aryl, heterocyclic aryl, amide and ester groups. An “aryl” group refers to an aromatic group which has at least one ring having a conjugated p electron system and includes carbocyclic aryl, heterocyclic aryl and biaryl groups, all of which may be optionally substituted. The preferred substituent(s) of aryl groups are halogen, trihalomethyl, hydroxyl, SH, OH, cyano, alkoxy, alkyl, alkenyl, alkynyl, and amino groups. An “alkylaryl” group refers to an alkyl group (as described above) covalently joined to an aryl group (as described above). Carbocyclic aryl groups are groups wherein the ring atoms on the aromatic ring are all carbon atoms. The carbon atoms are optionally substituted. Heterocyclic aryl groups are groups having from 1 to 3 heteroatoms as ring atoms in the aromatic ring and the remainder of the ring atoms are carbon atoms. Suitable heteroatoms include oxygen, sulfur, and nitrogen, and include furanyl, thienyl, pyridyl, pyrrolyl, N-lower alkyl pyrrolo, pyrimidyl, pyrazinyl, imidazolyl and the like, all optionally substituted. An “amide” refers to an —C(O)—NH—R, where R is either alkyl, aryl, alkylaryl or hydrogen. An “ester” refers to an —C(O)—OR′, where R is either alkyl, aryl, alkylaryl or hydrogen.  
     [0117] By “nucleotide” as used herein is as recognized in the art to include natural bases (standard), and modified bases well known in the art. Such bases are generally located at the 1′ position of a nucleotide sugar moiety. Nucleotides generally comprise a base, sugar and a phosphate group. The nucleotides can be unmodified or modified at the sugar, phosphate and/or base moiety, (also referred to interchangeably as nucleotide analogs, modified nucleotides, non-natural nucleotides, non-standard nucleotides and other; see for example, Usman and McSwiggen, supra; Eckstein et al., International PCT Publication No. WO 92/07065; Usman et al., International PCT Publication No. WO 93/15187; Uhlman &amp; Peyman, supra. All these publications are hereby incorporated by reference herein). There are several examples of modified nucleic acid bases known in the art as summarized by Limbach et al., 1994,  Nucleic Acids Res.  22, 2183. Some of the non-limiting examples of base modifications that can be introduced into nucleic acid molecules include, inosine, purine, pyridin-4-one, pyridin-2-one, phenyl, pseudouracil, 2,4,6-trimethoxy benzene, 3-methyl uracil, dihydrouridine, naphthyl, aminophenyl, 5-alkylcytidines (e.g., 5-methylcytidine), 5-alkyluridines (e.g., ribothymidine), 5-halouridine (e.g., 5-bromouridine) or 6-azapyrimidines or 6-alkylpyrimidines (e.g. 6-methyluridine), propyne, and others (Burgin et al., 1996,  Biochemistry,  35, 14090; Uhlman &amp; Peyman, supra). All these publications are incorporated by reference herein. By “modified bases” in this aspect is meant nucleotide bases other than adenine, guanine, cytosine and uracil at 1′ position or their equivalents; such bases may be used at any position, for example, within the catalytic core of an enzymatic nucleic acid molecule and/or in the substrate-binding regions of the nucleic acid molecule.  
     [0118] In a preferred embodiment, the invention features modified ribozymes with phosphate backbone modifications comprising one or more phosphorothioate, phosphorodithioate, methylphosphonate, morpholino, amidate carbamate, carboxymethyl, acetamidate, polyamide, sulfonate, sulfonamide, sulfamate, formacetal, thioformacetal, and/or alkylsilyl, substitutions. For a review of oligonucleotide backbone modifications see Hunziker and Leumann, 1995,  Nucleic Acid Analogues: Synthesis and Properties,  in  Modern Synthetic Methods,  VCH, 331-417, and Mesmaeker et al., 1994,  Novel Backbone Replacements for Oligonucleotides,  in  Carbohydrate Modifications in Antisense Research,  ACS, 24-39. These references are hereby incorporated by reference herein.  
     [0119] By “abasic” is meant sugar moieties lacking a base or having other chemical groups in place of a base at the 1′ position, (for more details, see Wincott et al., International PCT publication No. WO 97/26270).  
     [0120] By “unmodified nucleoside” is meant one of the bases adenine, cytosine, guanine, thymine, uracil joined to the 1′ carbon of β-D-ribo-furanose.  
     [0121] By “modified nucleoside” is meant any nucleotide base which contains a modification in the chemical structure of an unmodified nucleotide base, sugar and/or phosphate.  
     [0122] In connection with 2′-modified nucleotides as described for the present invention, by “amino” is meant 2′-NH 2  or 2′-O—NH 2 , which may be modified or unmodified. Such modified groups are described, for example, in Eckstein et al., U.S. Pat. No. 5,672,695 and Matulic-Adamic et al., WO 98/28317, respectively, which are both incorporated by reference in their entireties.  
     [0123] Various modifications to nucleic acid (e.g., antisense and ribozyme) structure can be made to enhance the utility of these molecules. Such modifications will enhance shelf-life, half-life in vitro, stability, and ease of introduction of such oligonucleotides to the target site, e.g., to enhance penetration of cellular membranes, and confer the ability to recognize and bind to targeted cells.  
     [0124] Use of these molecules will lead to better treatment of the disease progression by affording the possibility of combination therapies (e.g., multiple ribozymes targeted to different genes, ribozymes coupled with known small molecule inhibitors, or intermittent treatment with combinations of ribozymes (including different ribozyme motifs) and/or other chemical or biological molecules). The treatment of patients with nucleic acid molecules may also include combinations of different types of nucleic acid molecules. Therapies may be devised which include a mixture of ribozymes (including different ribozyme motifs), antisense and/or 2-5A chimera molecules to one or more targets to alleviate symptoms of a disease.  
     [0125] Administration of Ribozymes  
     [0126] Sullivan et al., PCT WO 94/02595, describes the general methods for delivery of enzymatic RNA molecules. Ribozymes may be administered to cells by a variety of methods known to those familiar to the art, including, but not restricted to, encapsulation in liposomes, by iontophoresis, or by incorporation into other vehicles, such as hydrogels, cyclodextrins, biodegradable nanocapsules, and bioadhesive microspheres. For some indications, ribozymes may be directly delivered ex vivo to cells or tissues with or without the aforementioned vehicles. Alternatively, the RNA/vehicle combination is locally delivered by direct injection or by use of a catheter, infusion pump, stent or other delivery devices such as Alzet® pumps, Medipad® devices. Other routes of delivery include, but are not limited to, intravascular, intramuscular, subcutaneous or joint injection, aerosol inhalation, oral (tablet or pill form), topical, systemic, ocular, intraperitoneal and/or intrathecal delivery. More detailed descriptions of ribozyme delivery and administration are provided in Sullivan et al., supra and Draper et al., PCT WO93/23569, which have been incorporated by reference herein.  
     [0127] The molecules of the instant invention can be used as pharmaceutical agents. Pharmaceutical agents prevent, inhibit the occurrence, or treat (alleviate a symptom to some extent, preferably all of the symptoms) of a disease state in a patient.  
     [0128] The negatively charged polynucleotides of the invention can be administered (e.g., RNA, DNA or protein) and introduced into a patient by any standard means, with or without stabilizers, buffers, and the like, to form a pharmaceutical composition. When it is desired to use a lipid or liposome delivery mechanism, standard protocols for formulation can be followed. The compositions of the present invention may also be formulated and used as tablets, capsules or elixirs for oral administration; suppositories for rectal administration; sterile solutions; suspensions for injectable administration; and the like.  
     [0129] The present invention also includes pharmaceutically acceptable formulations of the compounds described. These formulations include salts of the above compounds, e.g., acid addition salts, including salts of hydrochloric, hydrobromic, acetic acid, and benzene sulfonic acid.  
     [0130] A pharmacological composition or formulation refers to a composition or formulation in a form suitable for administration, e.g., systemic administration, into a cell or patient, preferably a human. Suitable forms, in part, depend upon the use or the route of entry, for example oral, transdermal, or by injection. Such forms should not prevent the composition or formulation to reach a target cell (i.e., a cell to which the negatively charged polymer is desired to be delivered to). For example, pharmacological compositions injected into the blood stream should be soluble. Other factors are known in the art, and include considerations such as toxicity and forms which prevent the composition or formulation from exerting its effect.  
     [0131] By “systemic administration” is meant in vivo systemic absorption or accumulation of drugs in the blood stream followed by distribution throughout the entire body. Administration routes which lead to systemic absorption include, without limitations: intravenous, subcutaneous, intraperitoneal, inhalation, oral, intrapulmonary and intramuscular. Each of these administration routes expose the desired negatively charged polymers, e.g., nucleic acids, to an accessible diseased tissue. The rate of entry of a drug into the circulation has been shown to be a function of molecular weight or size. The use of a liposome or other drug carrier comprising the compounds of the instant invention can potentially localize the drug, for example, in certain tissue types, such as the tissues of the reticular endothelial system (RES). A liposome formulation which can facilitate the association of drug with the surface of cells, such as, lymphocytes and macrophages is also useful. This approach may provide enhanced delivery of the drug to target cells by taking advantage of the specificity of macrophage and lymphocyte immune recognition of abnormal cells, such as the HCV infected liver cells.  
     [0132] The invention also features the use of a composition comprising surface-modified liposomes containing poly (ethylene glycol) lipids (PEG-modified, or long-circulating liposomes or stealth liposomes). These formulations offer a method for increasing the accumulation of drugs in target tissues. This class of drug carriers resists opsonization and elimination by the mononuclear phagocytic system (MPS or RES), thereby enabling longer blood circulation times and enhanced tissue exposure for the encapsulated drug (Lasic et al.  Chem. Rev.  1995, 95, 2601-2627; Ishiwata et al.,  Chem. Pharm. Bull.  1995, 43, 1005-1011). All these publications are incorporated by reference herein. Such liposomes have been shown to accumulate selectively in tumors, presumably by extravasation and capture in the neovascularized target tissues (Lasic et al.,  Science  1995, 267, 1275-1276; Oku et al.,1995,  Biochim. Biophys. Acta,  1238, 86-90). All these references are incorporated by reference herein. The long-circulating liposomes enhance the pharmacokinetics and pharmacodynamics of DNA and RNA, particularly compared to conventional cationic liposomes which are known to accumulate in tissues of the MPS (Liu et al.,  J. Biol. Chem.  1995, 42, 24864-24870; Choi et al., International PCT Publication No. WO 96/10391; Ansell et al., International PCT Publication No. WO 96/10390; Holland et al., International PCT Publication No. WO 96/10392; all of these are incorporated by reference herein).  
     [0133] In addition other cationic molecules may also be utilized to deliver the molecules of the present invention. For example, ribozymes may be conjugated to glycosylated poly(L-lysine) which has been shown to enhance localization of antisense oligonucleotides into the liver (Nakazono et al., 1996,  Hepatology  23, 1297-1303; Nahato et al., 1997,  Biochem Pharm.  53, 887-895). Glycosylated poly (L-lysine) may be covalently attached to the enzymatic nucleic acid or be bound to enzymatic nucleic acid through electrostatic interaction.  
     [0134] The present invention also includes compositions prepared for storage or administration which include a pharmaceutically effective amount of the desired compounds in a pharmaceutically acceptable carrier or diluent. Acceptable carriers or diluents for therapeutic use are well known in the pharmaceutical art, and are described, for example, in  Remington&#39;s Pharmaceutical Sciences,  Mack Publishing Co. (A. R. Gennaro edit. 1985) hereby incorporated by reference herein. For example, preservatives, stabilizers, dyes and flavoring agents may be provided. Id. at 1449. These include sodium benzoate, sorbic acid and esters of p-hydroxybenzoic acid. In addition, antioxidants and suspending agents may be used.  
     [0135] A pharmaceutically effective dose is that dose required to prevent, inhibit the occurrence, or treat (alleviate a symptom to some extent, preferably all of the symptoms) a disease state. The pharmaceutically effective dose depends on the type of disease, the composition used, the route of administration, the type of mammal being treated, the physical characteristics of the specific mammal under consideration, concurrent medication, and other factors which those skilled in the medical arts will recognize. Generally, an amount between 0.1 mg/kg and 100 mg/kg body weight/day of active ingredients is administered dependent upon potency of the negatively charged polymer.  
     [0136] The nucleic acid molecules of the present invention may also be administered to a patient in combination with other therapeutic compounds to increase the overall therapeutic effect. The use of multiple compounds to treat an indication may increase the beneficial effects while reducing the presence of side effects.  
     [0137] Alternatively, the enzymatic nucleic acid molecules of the instant invention can be expressed within cells from eukaryotic promoters (e.g., Izant and Weintraub, 1985  Science  229, 345; McGarry and Lindquist, 1986  Proc. Natl. Acad. Sci. USA  83, 399; Scanlon et al., 1991,  Proc. Natl. Acad. Sci. USA,  88, 10591-5; Kashani-Sabet et al., 1992  Antisense Res. Dev.,  2, 3-15; Dropulic et al., 1992  J. Virol,  66, 1432-41; Weerasinghe et al., 1991  J. Virol,  65, 5531-4; Ojwang et al., 1992  Proc. Natl. Acad. Sci. USA  89, 10802-6; Chen et al, 1992  Nucleic Acids Res.,  20, 4581-9; Sarver et al., 1990  Science  247, 1222-1225; Thompson et al., 1995  Nucleic Acids Res.  23, 2259; Good et al., 1997,  Gene Therapy,  4, 45; all of these references are hereby incorporated in their totalities by reference herein). Those skilled in the art realize that any nucleic acid can be expressed in eukaryotic cells from the appropriate DNA/RNA vector. The activity of such nucleic acids can be augmented by their release from the primary transcript by a ribozyme (Draper et al., PCT WO 93/23569, and Sullivan et al., PCT WO 94/02595; Ohkawa et al., 1992  Nucleic Acids Symp. Ser.,  27, 15-6; Taira et al., 1991,  Nucleic Acids Res.,  19, 5125-30; Ventura et al., 1993  Nucleic Acids Res.,  21, 3249-55; Chowrira et al., 1994  J. Biol. Chem.  269, 25856; all of these references are hereby incorporated in their totalities by reference herein).  
     [0138] In another aspect of the invention, enzymatic nucleic acid molecules that cleave target molecules are expressed from transcription units (see for example Couture et al., 1996,  TIG.,  12, 510) inserted into DNA or RNA vectors. The recombinant vectors are preferably DNA plasmids or viral vectors. Ribozyme expressing viral vectors could be constructed based on, but not limited to, adeno-associated virus, retrovirus, adenovirus, or alphavirus. Preferably, the recombinant vectors capable of expressing the ribozymes are delivered as described above, and persist in target cells. Alternatively, viral vectors may be used that provide for transient expression of ribozymes. Such vectors might be repeatedly administered as necessary. Once expressed, the ribozymes cleave the target mRNA. The active ribozyme contains an enzymatic center or core equivalent to those in the examples, and binding arms able to bind target nucleic acid molecules such that cleavage at the target site occurs. Other sequences may be present which do not interfere with such cleavage. Delivery of ribozyme expressing vectors could be systemic, such as by intravenous or intramuscular administration, by administration to target cells ex-planted from the patient followed by reintroduction into the patient, or by any other means that would allow for introduction into the desired target cell (for a review see Couture et al., 1996,  TIG.,  12, 510).  
     [0139] In one aspect the invention features, an expression vector comprising nucleic acid sequence encoding at least one of the nucleic acid catalyst of the instant invention is disclosed. The nucleic acid sequence encoding the nucleic acid catalyst of the instant invention is operable linked in a manner which allows expression of that nucleic acid molecule.  
     [0140] In another aspect the invention features an expression vector comprising: a) a transcription initiation region (e.g., eukaryotic pol I, II or III initiation region); b) a transcription termination region (e.g., eukaryotic pol I, II or III termination region); c) a nucleic acid sequence encoding at least one of the nucleic acid catalyst of the instant invention; and wherein said sequence is operably linked to said initiation region and said termination region, in a manner which allows expression and/or delivery of said nucleic acid molecule. The vector may optionally include an open reading frame (ORF) for a protein operably linked on the 5′ side or the 3′-side of the sequence encoding the nucleic acid catalyst of the invention; and/or an intron (intervening sequences).  
     [0141] Transcription of the nucleic acid molecule sequences are driven from a promoter for eukaryotic RNA polymerase I (pol I), RNA polymerase II (pol II), or RNA polymerase III (pol III). Transcripts from pol II or pol III promoters will be expressed at high levels in all cells; the levels of a given pol II promoter in a given cell type will depend on the nature of the gene regulatory sequences (enhancers, silencers, etc.) present nearby. Prokaryotic RNA polymerase promoters are also used, providing that the prokaryotic RNA polymerase enzyme is expressed in the appropriate cells (Elroy-Stein and Moss, 1990,  Proc. Natl. Acad. Sci. USA,  87, 6743-7; Gao and Huang 1993,  Nucleic Acids Res..,  21, 2867-72; Lieber et al., 1993,  Methods Enzymol.,  217, 47-66; Zhou et al., 1990,  Mol. Cell. Biol.,  10, 4529-37). All of these references are incorporated by reference herein. Several investigators have demonstrated that nucleic acid molecules, such as ribozymes expressed from such promoters can function in mammalian cells (e.g. Kashani-Sabet et al., 1992,  Antisense Res. Dev.,  2, 3-15; Ojwang et al., 1992,  Proc. Natl. Acad. Sci. USA,  89, 10802-6; Chen et al., 1992,  Nucleic Acids Res.,  20, 4581-9; Yu et al., 1993,  Proc. Natl. Acad. Sci. USA,  90, 6340-4; L&#39;Huillier et al., 1992,  EMBO J.,  11, 4411-8; Lisziewicz et al., 1993,  Proc. Natl. Acad. Sci. U.S.A,  90, 8000-4; Thompson et al, 1995,  Nucleic Acids Res.,  23, 2259; Sullenger &amp; Cech, 1993,  Science,  262, 1566). More specifically, transcription units such as the ones derived from genes encoding U6 small nuclear (snRNA), transfer RNA (tRNA) and adenovirus VA RNA are useful in generating high concentrations of desired RNA molecules such as ribozymes in cells (Thompson et al., supra; Couture and Stinchcomb, 1996, supra; Noonberg et al, 1994,  Nucleic Acid Res.,  22, 2830; Noonberg et al., U.S. Pat. No. 5,624,803; Good et al, 1997,  Gene Ther.,  4, 45; Beigelman et al., International PCT Publication No. WO 96/18736; all of these publications are incorporated by reference herein. The above ribozyme transcription units can be incorporated into a variety of vectors for introduction into mammalian cells, including but not restricted to, plasmid DNA vectors, viral DNA vectors (such as, adenovirus or adeno-associated virus vectors), or viral RNA vectors (such as retroviral or alphavirus vectors) (for a review, see Couture and Stinchcomb, 1996, supra).  
     [0142] In yet another aspect, the invention features an expression vector comprising nucleic acid sequence encoding at least one of the nucleic acid molecules of the invention, in a manner which allows expression of that nucleic acid molecule. The expression vector comprises in one embodiment; a) a transcription initiation region; b) a transcription termination region; c) a nucleic acid sequence encoding at least one said nucleic acid molecule; and wherein said sequence is operably linked to said initiation region and said termination region, in a manner which allows expression and/or delivery of said nucleic acid molecule. In another preferred embodiment the expression vector comprises: a) a transcription initiation region; b) a transcription termination region; c) an open reading frame; d) a nucleic acid sequence encoding at least one said nucleic acid molecule, wherein said sequence is operably linked to the 3′-end of said open reading frame; and wherein said sequence is operably linked to said initiation region, said open reading frame and said termination region, in a manner which allows expression and/or delivery of said nucleic acid molecule. In yet another embodiment, the expression vector comprises: a) a transcription initiation region; b) a transcription termination region; c) an intron; d) a nucleic acid sequence encoding at least one said nucleic acid molecule; and wherein said sequence is operably linked to said initiation region, said intron and said termination region, in a manner which allows expression and/or delivery of said nucleic acid molecule. In another embodiment, the expression vector comprises: a) a transcription initiation region; b) a transcription termination region; c) an intron; d) an open reading frame; e) a nucleic acid sequence encoding at least one said nucleic acid molecule, wherein said sequence is operably linked to the 3′-end of said open reading frame; and wherein said sequence is operably linked to said initiation region, said intron, said open reading frame and said termination region, in a manner which allows expression and/or delivery of said nucleic acid molecule.  
     [0143] Interferons  
     [0144] Type I interferons (IFN) are a class of natural cytokines that includes a family of greater than 25 IFN-α (Pesta, 1986,  Methods Enzymol.  119, 3-14) as well as IFN-β, and IFN-ω. Although evolutionarily derived from the same gene (Diaz et al., 1994,  Genomics  22, 540-552), there are many differences in the primary sequence of these molecules, implying an evolutionary divergence in biologic activity. All type I IFN share a common pattern of biologic effects that begin with binding of the IFN to the cell surface receptor (Pfeffer &amp; Strulovici, 1992, Transmembrane secondary messengers for IFN-α/β. In: Interferon.  Principles and Medical Applications.,  S. Baron, D. H. Coopenhaver, F. Dianzani, W. R. Fleischmann Jr., T. K. Hughes Jr., G. R. Kimpel, D. W. Niesel, G. J. Stanton, and S. K. Tyring, eds. 151-160). Binding is followed by activation of tyrosine kinases, including the Janus tyrosine kinases and the STAT proteins, which leads to the production of several IFN-stimulated gene products (Johnson et al., 1994,  Sci. Am.  270, 68-75). The IFN-stimulated gene products are responsible for the pleotropic biologic effects of type I IFN, including antiviral, antiproliferative, and immunomodulatory effects, cytokine induction, and HLA class I and class II regulation (Pestka et al., 1987,  Annu. Rev. Biochem  56, 727). Examples of IFN-stimulated gene products include 2-5-oligoadenylate synthetase (2-5 OAS), β 2 -microglobulin, neopterin, p 68  kinases, and the Mx protein (Chebath &amp; Revel, 1992, The 2-5 A system: 2-5 A synthetase, isospecies and functions. In: Interferon.  Principles and Medical Applications.  S. Baron, D. H. Coopenhaver, F. Dianzani, W. R. Jr. Fleischmann, T. K. Jr Hughes, G. R. Kimpel, D. W. Niesel, G. J. Stanton, and S. K. Tyring, eds., pp. 225-236; Samuel, 1992, The RNA-dependent P1/eIF-2α protein kinase. In:  Interferon. Principles and Medical Applications.  S. Baron, D. H. Coopenhaver, F. Dianzani, W. R. Fleischmann Jr., T. K. Hughes Jr., G. R. Kimpel, D. W. Niesel, G. H. Stanton, and S. K. Tyring, eds. 237-250; Horisberger, 1992, MX protein: function and Mechanism of Action. In: Interferon.  Principles and Medical Applications.  S. Baron, D. H. Coopenhaver, F. Dianzani, W. R. Fleischmann Jr., T. K. Hughes Jr., G. R. Kimpel, D. W. Niesel, G. H. Stanton, and S. K. Tyring, eds. 215-224). Although all type I IFN have similar biologic effects, not all the activities are shared by each type I IFN, and, in many cases, the extent of activity varies quite substantially for each IFN subtype (Fish et al, 1989,  J. Interferon Res.  9, 97-114; Ozes et al., 1992,  J. Interferon Res.  12, 55-59). More specifically, investigations into the properties of different subtypes of IFN-α, and molecular hybrids of IFN-α. have shown differences in pharmacological properties (Rubinstein, 1987,  J. Interferon Res.  7, 545-551). These pharmacological differences may arise from as few as three amino acid residue changes (Lee et al., 1982,  Cancer Res.  42, 1312-1316).  
     [0145] Eighty-five to 166 amino acids are conserved in the known IFN-α subtypes. Excluding the IFN-α pseudogenes, there are approximately 25 known distinct IFN-α subtypes. Pairwise comparisons of these nonallelic subtypes show primary sequence differences ranging from 2% to 23%. In addition to the naturally occurring IFNs, a non-natural recombinant type I interferon known as consensus interferon (CIFN) has been synthesized as a therapeutic compound (Tong et al., 1997,  Hepatology  26, 747-754).  
     [0146] Interferon is currently in use for at least 12 different indications including infectious and autoimmune diseases and cancer (Borden, 1992,  N. Engl. J. Med.  326, 1491-1492). For autoimmune diseases IFN has been utilized for treatment of rheumatoid arthritis, multiple sclerosis, and Crohn&#39;s disease. For treatment of cancer IFN has been used alone or in combination with a number of different compounds. Specific types of cancers for which IFN has been used include squamous cell carcinomas, melanomas, hypemephromas, hemangiomas, hairy cell leukemia, and Kaposi&#39;s sarcoma. In the treatment of infectious diseases, IFNs increase the phagocytic activity of macrophages and cytotoxicity of lymphocytes and inhibits the propagation of cellular pathogens. Specific indications for which IFN has been used as treatment include: hepatitis B, human papillomavirus types 6 and 11 (i.e. genital warts) (Leventhal et al., 1991,  N Engl J Med  325, 613-617), chronic granulomatous disease, and hepatitis C virus.  
     [0147] Numerous well controlled clinical trials using IFN-alpha in the treatment of chronic HCV infection have demonstrated that treatment three times a week results in lowering of serum ALT values in approximately 50% (range 40% to 70%) of patients by the end of 6 months of therapy (Davis et al., 1989,  New England Journal of Medicine  321, 1501-1506; Marcellin et al., 1991,  Hepatology  13, 393-397; Tong et al., 1997,  Hepatology  26, 747-754; Tong et al.,  Hepatology  26, 1640-1645). However, following cessation of interferon treatment, approximately 50% of the responding patients relapsed, resulting in a “durable” response rate as assessed by normalization of serum ALT concentrations of approximately 20 to 25%. In addition, studies that have examined six months of type 1 interferon therapy using changes in HCV RNA values as a clinical endpoint have demonstrated that up to 35% of patients will have a loss of HCV RNA by the end of therapy (Tong et al., 1997, supra). However, as with the ALT endpoint, about 50% of the patients relapse six months following cessation of therapy resulting in a durable virologic response of only 12% (23). Studies that have examined 48 weeks of therapy have demonstrated that the sustained virological response is up to 25%.  
     [0148] Ribozymes in combination with IFN have the potential to improve the effectiveness of treatment of HCV or any of the other indications discussed above. Ribozymes targeting RNAs associated with diseases such as infectious diseases, autoimmune diseases, and cancer, can be used individually or in combination with other therapies such as IFN to achieve enhanced efficacy.  
     EXAMPLES  
     [0149] The following are non-limiting examples showing the selection, isolation, synthesis and activity of enzymatic nucleic acids of the instant invention.  
     [0150] The following examples demonstrate the selection of ribozymes that cleave HCV RNA. The methods described herein represent a scheme by which ribozymes may be derived that cleave other RNA targets required for HCV replication.  
     Example 1  
     [0151] Identification of Potential Ribozyme Cleavage Sites in HCV RNA  
     [0152] The sequence of HCV RNA was screened for accessible sites using a computer folding algorithm. Regions of the mRNA that did not form secondary folding structures and contained potential hammerhead and/or hairpin ribozyme cleavage sites were identified. The sequences of these cleavage sites are shown in Tables IV-VIII, and X.  
     Example 2  
     [0153] Selection of Ribozyme Cleavage Sites in HCV RNA  
     [0154] To test whether the sites predicted by the computer-based RNA folding algorithm corresponded to accessible sites in HCV RNA, 20 hammerhead sites were selected for analysis. Ribozyme target sites were chosen by analyzing genomic sequences of HCV (Input Sequence=HPCJTA (Acc#D11168 &amp; D01171)) and prioritizing the sites on the basis of folding. Hammerhead ribozymes were designed that could bind each target (see FIG. 1) and were individually analyzed by computer folding (Christoffersen et al., 1994  J. Mol. Struc. Theochem,  311, 273; Jaeger et al., 1989,  Proc. Natl. Acad. Sci. USA,  86, 7706) to assess whether the ribozyme sequences fold into the appropriate secondary structure. Those ribozymes with unfavorable intramolecular interactions between the binding arms and the catalytic core were eliminated from consideration. As noted below, varying binding arm lengths can be chosen to optimize activity. Generally, at least 5 bases on each arm are able to bind to, or otherwise interact with, the target RNA.  
     [0155] Selection of ribozyme candidates was initiated by scanning for all hammerhead cleavage sites in an HCV RNA sequence derived from a patient infected with HCV genotype 1 b.  The results of this sequence analysis are shown in Table III. As seen by Table III, 1300 hammerhead ribozyme sites were identified by this analysis. Next, in order to identify hammerhead ribozyme candidates that would cleave in the conserved regions of the HCV genome, a sequence alignment of approximately 50 HCV isolates from genotypes 1a, 1b, 2a, 2b, 2c, 3a, 3b, 4a, 5a, and 6 was completed. Within genotype sites were identified that are in areas having the greatest sequence identity between all isolates examined. This analysis reduced the hammerhead ribozyme candidates to about 23 (Table III).  
     [0156] Due to the high sequence variability of the HCV genome, selection of ribozymes for broad therapeutic applications should probably involve the conserved regions of the HCV genome. A list of the thirty-hammerhead ribozymes derived from the conserved regions (5′-Non-Coding Region (NCR), 5′-end of core protein coding region, and 3′-NCR) of the HCV genome is shown in Table IV. In general, ribozymes targeted to sites located in the 5′ terminal region of the HCV genome should block translation while ribozymes cleavage sites located in the 3′ terminal region of the genome should block RNA replication.  
     Example 3  
     [0157] Chemical Synthesis and Purification of Ribozymes  
     [0158] Ribozymes of the hammerhead or hairpin motif were designed to anneal to various sites in the RNA message. The binding arms are complementary to the target site sequences described above. The ribozymes were chemically synthesized. The method of synthesis used followed the procedure for normal RNA synthesis as described in Usman et al., (1987  J. Am. Chem. Soc.,  109, 7845), Scaringe et al., (1990  Nucleic Acids Res.,  18, 5433) and Wincott et al., supra, and made use of common nucleic acid protecting and coupling groups, such as dimethoxytrityl at the 5′-end, and phosphoramidites at the 3′-end. The average stepwise coupling yields were &gt;98%.  
     [0159] Inactive hammerhead ribozymes were synthesized by substituting switching the order of G 5 A 6  and substituting a U for A 14  (numbering from Hertel et al., 1992  Nucleic Acids Res.,  20, 3252). Hairpin ribozymes were synthesized in two parts and annealed to reconstruct the active ribozyme (Chowrira and Burke, 1992  Nucleic Acids Res.,  20, 2835-2840). Ribozymes were also synthesized from DNA templates using bacteriophage T7 RNA polymerase (Milligan and Uhlenbeck, 1989,  Methods Enzymol.  180, 51). Ribozymes were modified to enhance stability by modification with nuclease resistant groups, for example, 2′-amino, 2′-C-allyl, 2′-flouro, 2′-O-methyl, 2′-H (for a review, see Usman and Cedergren, 1992  TIBS  17, 34). Ribozymes were purified by gel electrophoresis using general methods or were purified by high pressure liquid chromatography (HPLC; see Wincott et al., supra; the totality of which is hereby incorporated herein by reference) and were resuspended in water. The sequences of the chemically synthesized ribozymes used in this study are shown below in Tables IV-X.  
     Example 4  
     [0160] Ribozyme Cleavage of HCV RNA Target in vitro  
     [0161] Ribozymes targeted to the HCV are designed and synthesized as described above.  
     [0162] These ribozymes can be tested for cleavage activity in vitro, for example, using the following procedure. The target sequences and the nucleotide location within the HCV are given in Table IV.  
     [0163] Cleavage Reactions: Full-length or partially full-length, internally-labeled target RNA for ribozyme cleavage assay is prepared by in vitro transcription in the presence of [- 32 p] CTP, passed over a G 50Sephadex® column by spin chromatography and used as substrate RNA without further purification. Alternately, substrates are 5′ 32 p-end labeled using T4 polynucleotide kinase enzyme. Assays are performed by pre-warming a 2×concentration of purified ribozyme in ribozyme cleavage buffer (50 mM Tris-HCl, pH 7.5 at 37° C., 10 mM MgCl 2 ) and the cleavage reaction was initiated by adding the 2×ribozyme mix to an equal volume of substrate RNA (maximum of 1-5 nM) that was also pre-warmed in cleavage buffer. As an initial screen, assays are carried out for 1 hour at 37° C. using a final concentration of either 40 nM or 1 mM ribozyme, i.e., ribozyme excess. The reaction is quenched by the addition of an equal volume of 95% formamide, 20 mM EDTA, 0.05% bromophenol blue and 0.05% xylene cyanol after which the sample is heated to 95° C. for 2 minutes, quick chilled and loaded onto a denaturing polyacrylamide gel. Substrate RNA and the specific RNA cleavage products generated by ribozyme cleavage are visualized on an autoradiograph of the gel. The percentage of cleavage is determined by Phosphor Imager® quantitation of bands representing the intact substrate and the cleavage products.  
     Example 5  
     [0164] Ability of HCV Ribozymes to Cleave HCV RNA in Patient Serum  
     [0165] Ribozymes targeting sites in HCV RNA were synthesized using modifications that confer nuclease resistance (Beigelman, 1995,  J. Biol. Chem.  270, 25702). It has been well documented that serum from chronic hepatitis C patients contains on average 3×10 6  copies/ml of HCV RNA. To further select ribozyme product candidates, the 30 HCV specific ribozymes are characterized for HCV RNA cleavage activity utilizing HCV RNA isolated from the serum of genotype 1b HCV patients. The best candidates from the HCV genotype 1b screen will be screened against isolates from the wide range of HCV genotypes including 1a, 1b, 2a, 2b, 2c, 3a, 3b, 4a, 5a, and 6. Therefore, it is possible to select ribozyme candidates for further development based on their ability to broadly cleave HCV RNA from a diverse range of HCV genotypes and quasi-species.  
     Example 6  
     [0166] Ribozyme Cleavage of Conserved HCV RNA Target Sites in vitro  
     [0167] There are three regions of the genome that are highly conserved, both within a genotype and across different genotypes. These conserved sequences occur in the 5′ and 3′ non-coding regions (NCRs) as well as the 5′-end of the Core Protein coding region. These regions are thought to be important for HCV RNA replication and translation. Thus, therapeutic agents that target these conserved HCV genomic regions may have a significant impact over a wide range of HCV genotypes. The presence of quasi-species, and the potential for infection with more than one genotype makes this a critical feature of an effective therapy. Moreover, it is unlikely that drug resistance will occur, since mutations that have been suggested to lead to drug resistance typically do not occur within these highly conserved regions. In order to target multiple genotypes and decrease the chance of developing drug resistance, Applicant has designed ribozymes that cleave in regions of identity within the conserved regions discussed above.  
     [0168] Sequence alignments were performed for the 5′ NCR, the 5′ end of the Core Protein coding region, and the 3′ NCR. For the 5′ NCR, 34 different isolates representing genotypes 1a, 1b, 2a, 2b, 2c, 3a, 3b, 4a, 4f, and 5a were aligned. The alignments included the sequences from nucleotide position 1 to nucleotide position 350 (18 nucleotides downstream of the initiator ATG codon), using the reported sequence “HPCK1S1” as the reference for numbering. For the Core Protein coding region, 44 different isolates representing genotypes 1a, 1b, 2a, 2b, 2c, 3a, 3b, 4a, 4c, 4f, 5a, and 6a were aligned. These alignments included 600 nucleotides, beginning 8 nucleotides upstream of the initiator ATG codon. As the reference for numbering, the reported sequence “HPCCOPR” was used, with the “C” eight nucleotides upstream of the initiator codon ATG designated as “1”. For the 3′ NCR region, 20 different isolates representing genotypes 1b, 2a, 2b, 3a, and 3b were aligned. These alignments included sequences in the 3′ terminal 235 nucleotides of the genome, with the reported sequence “D85516” used as the reference for numbering, and the 235 th  nucleotide from the 3′ end designated as “1”.  
     [0169] During analysis of the alignments of each region, each sequence was compared to the respective reference sequence (identified above), and regions of identity across all isolates were determined. All potential ribozyme sites were identified in the reference sequence. The highest priority for choosing ribozyme sites was that the site should have 100% identity across all isolates aligned, at every position in both the cleavage site and binding arms. Ribozyme sites that met these criteria were chosen. In addition, two specific allowances were made as follows. 1) If a potential ribozyme site had 100% sequence identity at all except one or two nucleotide positions, then the actual nucleotide at that position was examined in the isolate(s) that differed. If that nucleotide was such that a ribozyme designed to allow “G:U wobble” base-paring could function on all the isolates, then that site was chosen. 2) If a potential ribozyme site had 100% sequence identity at all except one or two nucleotide positions, then the genotype of the isolate which contained the differing nucleotide(s) was examined. If the genotype of the isolate that differed was of extremely rare prevalence, then that site was also chosen.  
     [0170] Ribozyme sites identified and referred to below use the following nomenclature: “region of the genome in which the site exists” followed by “nucleotide position 5′ to the cleavage site” (according to the reference sequence and numbering described above). For example, a ribozyme cleavage site at nucleotide position 67 in the 5′ NCR is designated “5-67”, and a ribozyme cleavage site at position 48 in the core coding region is designated “c48”.  
     [0171] A number of these ribozymes were screened in an in vitro HCV cleavage assay to select appropriate ribozyme candidates for cell culture studies. The ribozymes selected for screening targeted the 5′ UTR region that is necessary for HCV translation. These sites are all conserved among the 8 major HCV genotypes and 18 subtypes, and have a high degree of homology in every HCV isolate that was used in the analysis described above. HCV RNA of four different genotypes (1b, 2a, 4, and 5) were isolated from human patients and the 5′ HCV UTR and 5′ core region were amplified using RT-PCR. Run-off transcripts of the 5′ HCV UTR region (˜750 nt transcripts) were prepared from the RT-PCR products, which contained a T7 promoter, using the T7 Megascript® transcription kit and the manufacturers protocol (Ambion, Inc.). Unincorporated nucleotides are removed by spin column filtration on Bio-Gel P-60 resin (Bio-Rad). The filtered transcript was 5′ end labeled with  32 P using Polynucleotide Kinase (Boehringer/Mannheim) and 150μCi/μl Gamma-32P-ATP (NEN) using the enzyme manufacturer&#39;s protocol. The kinased transcript is spin purified again to remove unincorporated Gamma-32P-ATP and gel purified on 5% polyacrylamide gel.  
     [0172] Ribozymes targeting various sites from Table IV were selected and tested on the 5′ HCV UTR transcript sequence to test the efficiency of RNA cleavage. 15 ribozymes were synthesized as previously described (Wincott et al., supra).  
     [0173] Assays were performed by pre-warming a 2×(2 μM) concentration of purified ribozyme in ribozyme cleavage buffer (50 mM TRIS pH 7.5, 10 mM MgCl 2 , 10 units RNase Inhibitor (Boehringer/Mannheim), 10 mM DTT, 0.5μg tRNA) and the cleavage reaction was initiated by adding the 2×ribozyme mix to an equal volume of substrate RNA (17.46 pmole final concentration) that was also pre-warmed in cleavage buffer. The assay was carried out for 24 hours at 37° C. using a final concentration of 1 μM ribozyme, i.e., ribozyme excess. The reaction was quenched by the addition of an equal volume of 95% formamide, 20 mM EDTA, 0.05% bromophenol blue and 0.05% xylene cyanol after which the sample is heated to 95° C. for 2 minutes, quick chilled and loaded onto a denaturing polyacrylamide gel. Substrate RNA and the specific RNA cleavage products generated by ribozyme cleavage are visualized on an autoradiograph of the gel. The percentage of cleavage is determined by Phosphor Imager® quantitation of bands representing the intact substrate and the cleavage products.  
     [0174] Observed cleavage fragment sizes from the gels are correlated to predicted fragment sizes by comparison to the RNA marker. The optical density of expected cleavage fragments are determined from the phosphorimage plates and ranked from highest density, indicating the most cleavage product, to lowest of each genotype of HCV transcript tested. The top 3 cleaving ribozymes (out of 15 ribozymes tested) are given ranking values of 5, the next 3 highest densities are given ranking values of 4, etc. for every genotype tested. The ranking values for each ribozyme are averaged between the genotypes tested. Individual and average ribozyme ranking values are graphed and compared. The results (FIG. 2) demonstrate that many of these tested ribozymes are able to give high levels of cleavage regardless of genotype. In particular, ribozymes targeting site HCV0.5-258, HCV0.5-294, HCV0.5-313 (Sakamoto et al.,  J. Clinical Investigation  1996 98(12):2720-2728), and HCV0.5-318 (Table IV) appear to demonstrate a consistent pattern of RNA cleavage  
     Example 7  
     [0175] Inhibition of Luciferase Activity Using HCV Targeting Ribozymes in OST7 Cells  
     [0176] The capability of ribozymes to inhibit HCV RNA intracellularly was tested using a dual reporter system that utilizes both firefly and Renilla luciferase (FIG. 3).  
     [0177] The ribozymes targeted to the 5′ HCV UTR region, which when cleaved, would prevent the translation of the transcript into luciferase. OST-7 cells were plated at 12,500 cells per well in black walled 96 well plates (Packard) in medium DMEM containing 10% fetal bovine serum, 1% pen/strep, and 1% L-glutamine and incubated at 37° C. overnight.  
     [0178] A plasmid containing T7 promoter expressing 5′ HCV UTR and firefly luciferase (T7C1-341 (Wang et al., 1993,  J. of Virol.  67, 3338-3344)) was mixed with a pRLSV 40  Renilla control plasmid (Promega Corporation) followed by ribozyme, and cationic lipid to make a 5×concentration of the reagents (T7C1-341 (4 μg/ml), pRLSV40 renilla luciferase control (6 μg/ml), ribozyme (250 nM), transfection reagent (28.5 μg/ml).  
     [0179] The complex mixture was incubated at 37° C. for 20 minutes. The media was removed from the cells and 120 μl of Opti-mem media was added to the well followed by 30 μl of the 5×complex mixture. 150 μl of Opti-mem was added to the wells holding the untreated cells. The complex mixture was incubated on OST-7 cells for 4 hours, lysed with passive lysis buffer (Promega Corporation) and luminescent signals were quantified using the Dual Luciferase Assay Kit using the manufacturer&#39;s protocol (Promega Corporation). The ribozyme sequences used are given in Table IV. The ribozymes used were of the hammerhead motif. The hammerhead ribozymes were chemically modified such that the ribozyme consists of ribose residues at five positions (see for example FIG. 7); position 4 has either 2′-C-allyl or 2′-amino modification; position 7 has either 2′-amino modification or 2-O-methyl modification; the remaining nucleotide positions contain 2′-O-methyl substitutions; four nucleotides at the 5′ terminus contains phosphorothioate substitutions. Additionally, the 3′ end of the ribozyme includes a 3′-3′ linked inverted abasic moiety (abasic deoxyribose; iH). The data (FIG. 4) is given as a ratio between the firefly and Renilla luciferase fluorescence. All of the ribozymes targeting 5′ HCV UTR were able to reduce firefly luciferase signal relative to renilla luciferase.  
     Example 9  
     [0180] Ribozyme Mediated Inhibition of Luciferase Activity Compared to Its Inactive Control in OST-7 Cells  
     [0181] The dual reporter system described above was utilized to determine the level of reduction of luciferase activity mediated by a ribozyme compared to its inactive control. Ribozymes, having the chemical composition described in the previous example, to sites HCV 313 and 318 (Table IV) and their inactive controls were synthesized as above. The inactive control has the same nucleotide base composition as the active ribozyme but the nucleotide sequence has been scrambled. The protocols utilized for tissue culture and the luciferase assay was exactly as given in Example 8 except the ribozyme concentration in the 5×complex mixture was 1 mM (final concentration on the cells was 200 nM).  
     [0182] The results are given in FIG. 5. The ribozyme targeting HCV.5-318 was able to greatly reduce firefly luciferase activity compared to the untreated and inactive controls. The ribozyme targeting HCV.5-313 was able to slightly reduce firefly luciferase activity compared to the inactive control.  
     Example 10  
     [0183] Ribozyme Inhibition of Viral Replication  
     [0184] During HCV infection, viral RNA is present as a potential target for ribozyme cleavage at several processes: uncoating, translation, RNA replication and packaging. Target RNA may be more or less accessible to ribozyme cleavage at any one of these steps. Although the association between the HCV initial ribosome entry site (IRES) and the translation apparatus is mimicked in the HCV 5′UTR/luciferase reporter system (Example 9), these other viral processes are not represented in the OST7 system. The resulting RNA/protein complexes associated with the target viral RNA are also absent. Moreover, these processes may be coupled in an HCV-infected cell which could further impact target RNA accessibility. Therefore, we tested whether ribozymes designed to cleave the HCV 5′UTR could effect a replicating viral system.  
     [0185] Recently, Lu and Wimmer characterized an HCV-poliovirus chimera in which the poliovirus IRES was replaced by the IRES from HCV (Lu &amp; Wimmer, 1996,  Proc. Nat. Acad. Sci. USA.  93, 1412-1417). Poliovirus (PV) is a positive strand RNA virus like HCV, but unlike HCV is non-enveloped and replicates efficiently in cell culture. The HCV-PV chimera expresses a stable, small plaque phenotype relative to wild type PV.  
     [0186] The following ribozymes were synthesized for the experiment (Table VIII): ribozyme targeting site 183 (3 5′-end phosphorothioate linkages), scrambled control to site 183, ribozyme to site 318 (3 5′-end phosphorothioate linkages), ribozyme targeting site 183 (4 5′-end phosphorothioate linkages), inactive ribozyme targeting site 183 (4 5′-end phosphorothioate linkages). HeLa cells were infected with the HCV-PV chimera for 30 minutes and immediately treated with ribozyme. HeLa cells were seeded in U-bottom 96-well plates at a density of 9000-10,000 cells/well and incubated at 37° C. under 5% CO 2  for 24 h. Transfection of ribozyme (200 nM) was achieved by mixing of 10×ribozyme (2000 nM) and 10×of a cationic lipid (80 μg/ml) in DMEM (Gibco BRL) with 5% fetal bovine serum (FBS). Ribozyme/lipid complexes were allowed to incubate for 15 minutes at 37° C. under 5% CO 2 . Medium was aspirated from cells and replaced with 80 μls of DMEM (Gibco BRL) with 5% FBS serum, followed by the addition of 20 μls of 10×complexes. Cells were incubated with complexes for 24 hours at 37° C. under 5% CO 2 .  
     [0187] The yield of HCV-PV from treated cells (FIG. 6A) was quantified by plaque assay. The plaque assays were performed by diluting virus samples in serum-free DMEM (Gibco BRL) and applying 100 μl to HeLa cell monolayers (˜80% confluent) in 6-well plates for 30 minutes. Infected monolayers were overlayed with 3 ml 1.2% agar (Sigma) and incubated at 37° C. under 5% CO 2 . Two-three days later the overlay was removed, monolayers were stained with 1.2% crystal violet, and plaque forming units were counted. The data is shown in FIG. 6A. Ribozymes to site 183 inhibited HCV-PV replication by &gt;80% (P&lt;0.05) compared to the scrambled control (FIG. 6A, first two bars). In addition, 3 or 4 phosphorothioate stabilization was equally effective (P&lt;0.05 vs. control for each) in inhibiting viral replication (compare 1 st  and 4 th  bar in FIG. 6A). Ribozymes to the  318  site also had a statistically significant (P&lt;0.05), effect on viral replication (compare 2 nd  and  3   rd  bar in FIG. 6A).  
     [0188] To confirm that a ribozyme cleavage mechanism was responsible for the inhibition of HCV-PV replication observed, HCV-PV infected cells were treated with ribozymes to site 183 that maintained binding arm sequences but contained a mutation in the catalytic core to attenuate cleavage activity (Table I). Viral replication in these cells was not inhibited compared to cells treated with the scrambled control ribozyme (FIG. 6A, 4 th  and 5 th  bar), indicating that ribozyme cleavage activity was required for the inhibition of HCV-PV replication observed. In addition, ribozymes targeting site 183 of the HCV 5′UTR had no effect on wild type PV replication (FIG. 6B). These data provide evidence that the ribozyme-mediated inhibition of HCV-PV replication was dependent upon the HCV 5′UTR and not a general inhibition of PV replication.  
     [0189] Ribozymes to site 183 were also tested for the ability to inhibit HCV-PV replication during a single infectious cycle in HeLa cells (FIG. 8). Cells treated with ribozyme to site 183 (7/4 format) produced significantly less virus than cells treated with the scrambled control (&gt;80% inhibition at 8h post infection, P&lt;0.001).  
     Example 11  
     [0190] Shortening of Ribozyme Lengths  
     [0191] All the ribozymes described in example 10 above contained 7 nucleotides on each binding arms and contained a 4 base-paired stem II element (7/4 format). For pharmaceutical manufacture of a therapeutic ribozyme it is advantageous to minimize sequence length if possible. Thus ribozymes to site 183 were shortened by removing the outer most nucleotide from each binding arm such that the ribozyme has six nucleotides in each binding arm and the stem II region is four base-paired long (6/4 format); removing one base-pair (2 nucleotides) in stem II resulting in a 3 base-paired stem II (7/3 format); or removing one nucleotide from each binding arm and shortening the stem II by one base-pair (6/3 format). (See FIG. 7 for a schematic representation of each of these ribozymes). Ribozymes in all tested formats gave significant inhibition of viral replication (FIG. 8) with the 7/4, 7/3 and 6/3 formats being almost identical at the 8 h timepoint (P&lt;0.001 across time course for all formats). The shortest ribozyme tested (6/3 format) was slightly more efficacious (&gt;90% inhibition, P&lt;0.001) than the 7/4 ribozyme (˜80% inhibition, P&lt;0.001). The 6/3 ribozyme may have a greater ability to access site 183 in the HCV-PV chimera.  
     Example 12  
     [0192] Combination Therapy of HCV Ribozymes and Interferon  
     [0193] HeLa cells (10,000 cells per well) were pre-treated with 12.5 Units/ml of Interferon alpha in complete media (DMEM+5% FBS) or pre-treated with complete media alone for 4 hours and then infected with HCV-PV at an MOI=0.1 for 30 minutes. The viral inoculum was then removed and 200 nM ribozyme targeted to HCV site 183 (Rz) or binding attenuated control, which has mutations in the catalytic core of the ribozyme that severely attenuates the activity of the ribozyme, (BAC) was delivered using cationic lipid in complete media for 24 hours. After 24 hours, the cells were lysed three times by freeze/thaw to release virus and virus was quantified by plaque assay. Viral yield is shown as mean plaque forming units per ml (pfu/ml)+SEM. The data is shown in FIG. 10.  
     [0194] Pre-treatment with interferon (IFN) reduces the viral yield by ˜10 −1  in control treated cells (BAC+IFN versus BAC). Ribozyme treated cells produce 2×10 −1  less virus than control-treated cells (Rz versus BAC). The combination of Rz and IFN treatment results in a synergistic 4×10 −2  reduction in viral yield (Rz+IFN versus BAC). An additive effect would result in only a 3×10 −1  reduction (1×10 −1 +2×10 −1 ).  
     Example 13  
     [0195] Inhibition of Hepatitis C Virus Using Various Ribozyme Motifs  
     [0196] A number of varying ribozyme motifs (RPI motifs I-III; FIG. 9), were tested for their ability to inhibit HCV propagation in tissue culture. An example of RPI motif I (G-cleaver) is described in Kore et al., 1998,  Nucleic Acids Research  26, 4116-4120, while an example of RPI motif II (Inozyme) is described in Ludwig &amp; Sproat, International PCT Publication No. WO 98/58058). RPI motif III is a new ribozyme motif which applicant has recently developed and an example of this motif was tested herein.  
     [0197] OST7 cells were maintained in Dulbecco&#39;s modified Eagle&#39;s medium (GIBCO BRL) supplemented with 10% fetal calf serum, L-glutamine (2 mM) and penicillin/streptomycin. For transfections, OST7 cells were seeded in black-walled 96-well plates (Packard Instruments) at a density of 12,500 cells/well and incubated at 37° C. under 5% CO 2  for 24 hours. Co-transfection of target reporter HCVT7 C (0.8 g/ml), control reporter pRLSV 40 , (1.2 μg/ml) and ribozyme, 50-200 nM was achieved by the following method: a 5×mixture of HCVT7 C (4 μg/ml), pRLSV40 (6 μg/ml), ribozyme (250-1000 nM) and cationic lipid (28.5 μg/ml) was made in 150 μls of OPTI-MEM (GIBCO BRL) minus serum. Reporter/ribozyme/lipid complexes were allowed to form for 20 minutes at 37° C. under 5% CO 2 . Medium was aspirated from OST7 cells and replaced with 120 μls of OPTI-MEM (GIBCO BRL) minus serum, immediately followed by the addition of 30 μls of 5×reporter/ribozyme/lipid complexes. Cells were incubated with complexes for 4 hours at 37° C. under 5% CO 2  . Luciferase assay was performed as described in example 7. The data is summarized in Table IX, with each motif&#39;s results listed along with its control. All of the ribozyme motifs were able to reduce the amount of HCV produced by the cells compared to the ribozymes not targeted to any HCV (irrelevant controls).  
     Example 14  
     [0198] General Protocol for Virus Infection and Ribozyme Delivery  
     [0199] HeLa cells were seeded in 96-well plates at a density of 9000-10,000 cells/well and incubated at 37° C. under 5% CO 2  for 24 h. Cells were infected with HCV-PV at an MOI−0.1 for 30 min. Transfection of ribozyme or control oligonucleotides (200 nM final) was achieved by mixing of 5×ribozyme or control oligonucleotides (1000 nM) and 5×cationic lipid (40 μg/ml at 5×, 800 ng/well final) in DMEM with 5% fetal bovine serum (FBS) in U-bottom 96-well plates. Ribozyme/lipid complexes were allowed to incubate for 15 min at 37° C. under 5% CO 2 . Medium was aspirated from cells and replaced with 80 μl of DMEM with 5% FBS serum, followed by the addition of 20 μl of 5×complexes. Cells were incubated with complexes for 24 h at 37° C. under 5% CO 2 . After 24 h cells were lysed by three freeze/thaw cycles to release virus and virus was quantified by plaque assay.  
     Example 15  
     [0200] General Protocol for HCV Plaque Assay  
     [0201] Virus samples were diluted in serum-free DMEM and 100 μl applied to HeLa cell monolayers (˜80% confluent) in 6-well plates for 30 min. Infected monolayers were overlayed with 3 ml 1.2% agar (Sigma, St. Louis, Mo.) and incubated at 37° C. under 5% CO 2 . When plaques were visible (after two to three days) the overlay was removed, monolayers were stained with 1.2% crystal violet, and plaque forming units were counted.  
     Example 16  
     [0202] Inhibition of Hepatitis C Virus Using Other Ribozyme Directed Against the HCV Minus Strand  
     [0203] HeLa cells in 96-well plates were infected with a chimeric Hepatitis C-Poliovirus (HCV-PV) at a multiplicity of infection (MOI) of 0.1. Virus inoculum was then replaced with media containing 5% serum and 200 nM ribozyme (Table X) or scrambled attenuated control (SAC), as indicated, complexed to cationic lipid. After 24 h cells were lysed 3 times by freeze/thaw and virus was quantified by plaque assay. Results are summarized in FIGS. 12 and 13. Plaque forming units (pfu)/ml are shown as the mean of triplicate samples+S.D.  
     Example 17  
     [0204] Dose Response of Ribozyme Directed Against the HCV Minus Strand  
     [0205] Cells were infected and treated with ribozyme as described in Example 16 except that various amounts (as indicated) of anti-HCV ribozyme RPI.15006 was mixed with a control oligonucleotide (SAC) to maintain a constant 200 nM total dose of nucleic acid for delivery. FIG. 14 shows the results of this study that demonstrates an effective dose response in cells to treatment with a ribozyme directed against the HCV minus strand.  
     Example 18  
     [0206] Dose Response of Ribozyme Directed Against the HCV Plus Strand Combined with Ribozymes Targeting the HCV Minus Strand  
     [0207] Cells were infected and treated with ribozyme as described in Example 16 except that various amounts (as indicated) of anti-HCV ribozyme RPI.13919, targeting the plus strand, was mixed with ribozymes targeting the minus strand, as noted, to maintain a constant 200 nM total dose of nucleic acid for delivery. FIG. 15 shows the results of this study that demonstrates an effective dose response in cells to treatment with a ribozyme (RPI 13919) directed against the HCV plus strand combined with a ribozyme targeting the HCV minus strand (RPI 14975).  
     Example 19  
     [0208] Inhibition of HCV in vivo  
     [0209] Ribozyme directed reduction of HCV in vivo was examined in a mouse model, generally described in Vierling, International PCT Publication No. WO 99/16307, using HCV RNA as an endpoint. The study compared mice treated with ribozymes compared to scrambled-attenuated-core ribozymes (SAC) and saline controls. Active ribozyme and SAC were dosed from day 5 through 20 post-transplant. Various modes of analysis were used including ANOVA of raw quantitative HCV RNA, Dunnett&#39;s of raw quantitative HCV RNA, ANOVA of log10 quantitative HCV RNA, Dunnett&#39;s of log 10  quantitative HCV RNA, and Chi Square of qualitative results (HCV RNA +/−). Treatment with active ribozyme (RPI 13918), resulted in significant reduction of HCV RNA at 12 and 16 days using quantitative analysis (p&lt;0.05 by Dunnett&#39;s using the log 10  transformed HCV RNA results for all observations). The use of qualitative assessment, by converting the quantitative data into positive or negative results, confirmed with same trend. This study suggests that treatment with active anti-HCV ribozymes results in a significant reduction in HCV RNA in a trimeric mouse model.  
     [0210] Cell Culture Assays  
     [0211] Although there have been reports of replication of HCV in cell culture (see below), these systems are difficult to replicate and have proven unreliable. Therefore, as was the case for development of other anti-HCV therapeutics such as interferon and ribavirin, after demonstration of safety in animal studies applicant can proceed directly into a clinical feasibility study.  
     [0212] Several recent reports have documented in vitro growth of HCV in human cell lines (Mizutani et al.,  Biochem Biophys Res Commun  1996 227(3):822-826; Tagawa et al.,  Journal of Gasteroenterology and Hepatology  1995 10(5):523-527; Cribier et al.,  Journal of General Virology  76(10):2485-2491; Seipp et al.,  Journal of General Virology  1997 78(10)2467-2478; lacovacci et al.,  Research Virology  1997 148(2):147-151; Iocavacci et al.,  Hepatology  1997 26(5) 1328-1337; Ito et al.,  Journal of General Virology  1996 77(5):1043-1054; Nakajima et al,  Journal of Virology  1996 70(5):3325-3329; Mizutani et al.,  Journal of Virology  1996 70(10):7219-7223; Valli et al.,  Res Virol  1995 146(4): 285-288; Kato et al.,  Biochem Biophys Res Comm  1995 206(3):863-869). Replication of HCV has been demonstrated in both T and B cell lines as well as cell lines derived from human hepatocytes. Demonstration of replication was documented using either RT-PCR based assays or the b-DNA assay. It is important to note that the most recent publications regarding HCV cell cultures document replication for up to 6-months.  
     [0213] In addition to cell lines that can be infected with HCV, several groups have reported the successful transformation of cell lines with cDNA clones of full-length or partial HCV genomes (Harada et al.,  Journal of General Virology  1995 76(5)1215-1221; Haramatsu et al.,  Journal of Viral Hepatitis  1997 4S(1):61-67; Dash et al.,  American Journal of Pathology  1997 151(2):363-373; Mizuno et al.,  Gasteroenterology  1995 109(6):1933-40; Yoo et al.,  Journal Of Virology  1995 69(1):32-38).  
     [0214] Animal Models  
     [0215] The best characterized animal system for HCV infection is the chimpanzee.  
     [0216] Moreover, the chronic hepatitis that results from HCV infection in chimpanzees and humans is very similar. Although clinically relevant, the chimpanzee model suffers from several practical impediments that make use of this model difficult. These include; high cost, long incubation requirements and lack of sufficient quantities of animals. Due to these factors, a number of groups have attempted to develop rodent models of chronic hepatitis C infection. While direct infection has not been possible several groups have reported on the stable transfection of either portions or entire HCV genomes into rodents (Yamamoto et al.,  Hepatology  1995 22(3): 847-855; Galun et al.,  Journal of Infectious Disease  1995 172(1):25-30; Koike et al.,  Journal of General Virology  1995 76(12)3031-3038; Pasquinelli et al.,  Hepatology  1997 25(3): 719-727; Hayashi et al.,  Princess Takamatsu Symp  1995 25:1430149; Mariya K, Yotsuyanagi H, Shintani Y, Fujie H, Ishibashi K, Matsuura Y, Miyamura T, Koike K. Hepatitis C virus core protein induces hepatic steatosis in transgenic mice.  Journal of General Virology  1997 78(7) 1527-1531; Takehara et al.,  Hepatology  1995 21(3):746-751; Kawamura et al.,  Hepatology  1997 25(4): 1014-1021). In addition, transplantation of HCV infected human liver into immunocompromised mice results in prolonged detection of HCV RNA in the animal&#39;s blood.  
     [0217] Vierling, International PCT Publication No. WO 99/16307, describes a method for expressing hepatitis C virus in an in vivo animal model. Viable, HCV infected human hepatocytes are transplanted into a liver parenchyma of a scid/scid mouse host.  
     [0218] The scid/scid mouse host is then maintained in a viable state, whereby viable, morphologically intact human hepatocytes persist in the donor tissue and hepatitis C virus is replicated in the persisting human hepatocytes. This model provides an effective means for the study of HCV inhibition by ribozymes in vivo.  
     [0219] Diagnostic Uses  
     [0220] Ribozymes of this invention may be used as diagnostic tools to examine genetic drift and mutations within diseased cells or to detect the presence of HCV RNA in a cell. The close relationship between ribozyme activity and the structure of the target RNA allows the detection of mutations in any region of the molecule, which alters the base-pairing and three-dimensional structure of the target RNA. By using multiple ribozymes described in this invention, one may map nucleotide changes, which are important to RNA structure and function in vitro, as well as in cells and tissues. Cleavage of target RNAs with ribozymes may be used to inhibit gene expression and define the role (essentially) of specified gene products in the progression of disease. In this manner, other genetic targets may be defined as important mediators of the disease. These experiments will lead to better treatment of the disease progression by affording the possibility of combination therapies (e.g., multiple ribozymes targeted to different genes, ribozymes coupled with known small molecule inhibitors, or intermittent treatment with combinations of ribozymes and/or other chemical or biological molecules). Other in vitro uses of ribozymes of this invention are well known in the art, and include detection of the presence of mRNAs associated with HCV related condition. Such RNA is detected by determining the presence of a cleavage product after treatment with a ribozyme using standard methodology.  
     [0221] In a specific example, ribozymes which can cleave only wild-type or mutant forms of the target RNA are used for the assay. The first ribozyme is used to identify wild-type RNA present in the sample and the second ribozyme can be used to identify mutant RNA in the sample. As reaction controls, synthetic substrates of both wild-type and mutant RNA can be cleaved by both ribozymes to demonstrate the relative ribozyme efficiencies in the reactions and the absence of cleavage of the “non-targeted” RNA species. The cleavage products from the synthetic substrates can also serve to generate size markers for the analysis of wild-type and mutant RNAs in the sample population. Thus each analysis can involve two ribozymes, two substrates and one unknown sample which will be combined into six reactions. The presence of cleavage products can be determined using an RNase protection assay so that full-length and cleavage fragments of each RNA can be analyzed in one lane of a polyacrylamide gel. It is not absolutely required to quantify the results to gain insight into the expression of mutant RNAs and putative risk of the desired phenotypic changes in target cells. The expression of mRNA whose protein product is implicated in the development of the phenotype (i.e., HCV) is adequate to establish risk. If probes of comparable specific activity are used for both transcripts, then a qualitative comparison of RNA levels will be adequate and will decrease the cost of the initial diagnosis. Higher mutant form to wild-type ratios will be correlated with higher risk whether RNA levels are compared qualitatively or quantitatively.  
     [0222] Additional Uses  
     [0223] Potential usefulness of sequence-specific enzymatic nucleic acid molecules of the instant invention might have many of the same applications for the study of RNA that DNA restriction endonucleases have for the study of DNA (Nathans et al., 1975  Ann. Rev. Biochem.  44:273). For example, the pattern of restriction fragments could be used to establish sequence relationships between two related RNAs, and large RNAs could be specifically cleaved to fragments of a size more useful for study. The ability to engineer sequence specificity of the enzymatic nucleic acid molecule is ideal for cleavage of RNAs of unknown sequence. Applicant describes the use of nucleic acid molecules to down-regulate gene expression of target genes in bacterial, microbial, fungal, viral, and eukaryotic systems including plant, or mammalian cells.  
     [0224] All patents and publications mentioned in the specification are indicative of the levels of skill of those skilled in the art to which the invention pertains. All references cited in this disclosure are incorporated by reference to the same extent as if each reference had been incorporated by reference in its entirety individually.  
     [0225] One skilled in the art would readily appreciate that the present invention is well adapted to carry out the objects and obtain the ends and advantages mentioned, as well as those inherent therein. The methods and compositions described herein as presently representative of preferred embodiments are exemplary and are not intended as limitations on the scope of the invention. Changes therein and other uses will occur to those skilled in the art, which are encompassed within the spirit of the invention, are defined by the scope of the claims.  
     [0226] It will be readily apparent to one skilled in the art that varying substitutions and modifications may be made to the invention disclosed herein without departing from the scope and spirit of the invention. Thus, such additional embodiments are within the scope of the present invention and the following claims.  
     [0227] The invention illustratively described herein suitably may be practiced in the absence of any element or elements, limitation or limitations which is not specifically disclosed herein. Thus, for example, in each instance herein any of the terms “comprising”, “consisting essentially of” and “consisting of” may be replaced with either of the other two terms. The terms and expressions which have been employed are used as terms of description and not of limitation, and there is no intention that in the use of such terms and expressions of excluding any equivalents of the features shown and described or portions thereof, but it is recognized that various modifications are possible within the scope of the invention claimed. Thus, it should be understood that although the present invention has been specifically disclosed by preferred embodiments, optional features, modification and variation of the concepts herein disclosed may be resorted to by those skilled in the art, and that such modifications and variations are considered to be within the scope of this invention as defined by the description and the appended claims.  
     [0228] In addition, where features or aspects of the invention are described in terms of Markush groups or other grouping of alternatives, those skilled in the art will recognize that the invention is also thereby described in terms of any individual member or subgroup of members of the Markush group or other group.  
     [0229] Other embodiments are within the following claims.  
     TABLE I  
     [0230] Characteristics of Naturally Occurring Ribozymes  
     [0231] Group I Introns  
     [0232] Size: ˜150 to &gt;1000 nucleotides.  
     [0233] Requires a U in the target sequence immediately 5′ of the cleavage site.  
     [0234] Binds 4-6 nucleotides at the 5′-side of the cleavage site.  
     [0235] Reaction mechanism: attack by the 3′-OH of guanosine to generate cleavage products with 3′-OH and 5′-guanosine.  
     [0236] Additional protein cofactors required in some cases to help folding and maintainance of the active structure.  
     [0237] Over 300 known members of this class. Found as an intervening sequence in  Tetrahymena thermophila  rRNA, fungal mitochondria, chloroplasts, phage T4, blue-green algae, and others.  
     [0238] Major structural features largely established through phylogenetic comparisons, mutagenesis, and biochemical studies [ i , i ].  
     [0239] Complete kinetic framework established for one ribozyme [ iii , iv , v , vi ].  
     [0240] Studies of ribozyme folding and substrate docking underway [ vii , viii , ix ].  
     [0241] Chemical modification investigation of important residues well established [ x , xi ].  
     [0242] The small (4-6 nt) binding site may make this ribozyme too non-specific for targeted RNA cleavage, however, the Tetrahymena group I intron has been used to repair a “defective” -galactosidase message by the ligation of new -galactosidase sequences onto the defective message [ xii ].  
     [0243] RNAse P RNA (M1 RNA)  
     [0244] Size: ˜290 to 400 nucleotides.  
     [0245] RNA portion of a ubiquitous ribonucleoprotein enzyme.  
     [0246] Cleaves tRNA precursors to form mature tRNA [ xiii ].  
     [0247] Reaction mechanism: possible attack by M 2+ -OH to generate cleavage products with 3′-OH and 5′-phosphate.  
     [0248] RNAse P is found throughout the prokaryotes and eukaryotes. The RNA subunit has been sequenced from bacteria, yeast, rodents, and primates.  
     [0249] Recruitment of endogenous RNAse P for therapeutic applications is possible through hybridization of an External Guide Sequence (EGS) to the target RNA [ xiv , xv ] 
     [0250] Important phosphate and 2′ OH contacts recently identified [ xvi , xvii ] 
     [0251] Group II Introns  
     [0252] Size: &gt;1000 nucleotides.  
     [0253] Trans cleavage of target RNAs recently demonstrated [ xviii , xix ] 
     [0254] Sequence requirements not fully determined.  
     [0255] Reaction mechanism: 2′-OH of an internal adenosine generates cleavage products with 3′-OH and a “lariat” RNA containing a 3′-5′ and a 2′-5′ branch point.  
     [0256] Only natural ribozyme with demonstrated participation in DNA cleavage [ xx , xxi ] in addition to RNA cleavage and ligation.  
     [0257] Major structural features largely established through phylogenetic comparisons [ xxii ].  
     [0258] Important 2′ OH contacts beginning to be identified [ xxiii ] 
     [0259] Kinetic framework under development [ xxiv ] 
     [0260] Neurospora VS RNA  
     [0261] Size: ˜144 nucleotides.  
     [0262] Trans cleavage of hairpin target RNAs recently demonstrated [ xxv ].  
     [0263] Sequence requirements not fully determined.  
     [0264] Reaction mechanism: attack by 2′-OH 5′ to the scissile bond to generate cleavage products with 2′,3′-cyclic phosphate and 5′-OH ends.  
     [0265] Binding sites and structural requirements not fully determined.  
     [0266] Only 1 known member of this class. Found in Neurospora VS RNA.  
     [0267] Hammerhead Ribozyme  
     [0268] (see text for references)  
     [0269] Size: ˜13 to 40 nucleotides.  
     [0270] Requires the target sequence UH immediately 5′ of the cleavage site.  
     [0271] Binds a variable number nucleotides on both sides of the cleavage site.  
     [0272] Reaction mechanism: attack by 2′-OH 5′ to the scissile bond to generate cleavage products with 2′,3′-cyclic phosphate and 5′-OH ends.  
     [0273] 14 known members of this class. Found in a number of plant pathogens (virusoids) that use RNA as the infectious agent.  
     [0274] Essential structural features largely defined, including 2 crystal structures [ xxvi , xxvii ] 
     [0275] Minimal ligation activity demonstrated (for engineering through in vitro selection) [ xxviii ] 
     [0276] Complete kinetic framework established for two or more ribozymes [ xxix ] 
     [0277] Chemical modification investigation of important residues well established [ xxx ].  
     [0278] Hairpin Ribozyme  
     [0279] Size: ˜50 nucleotides.  
     [0280] Requires the target sequence GUC immediately 3′ of the cleavage site.  
     [0281] Binds 4-6 nucleotides at the 5′-side of the cleavage site and a variable number to the 3′-side of the cleavage site.  
     [0282] Reaction mechanism: attack by 2′-OH 5′ to the scissile bond to generate cleavage products with 2 ′,3′-cyclic phosphate and 5′-OH ends.  
     [0283] 3 known members of this class. Found in three plant pathogen (satellite RNAs of the tobacco ringspot virus, arabis mosaic virus and chicory yellow mottle virus) which uses RNA as the infectious agent.  
     [0284] Essential structural features largely defined [ xxxi , xxxii , xxxiii , xxxiv ] 
     [0285] Ligation activity (in addition to cleavage activity) makes ribozyme amenable to engineering through in vitro selection [ xxxv ] 
     [0286] Complete kinetic framework established for one ribozyme [ xxxvi ].  
     [0287] Chemical modification investigation of important residues begun [ xxxvii , xxxviii ] 
     [0288] Hepatitis Delta Virus (HDV) Ribozym  
     [0289] Size: ˜60 nucleotides.  
     [0290] Trans cleavage of target RNAs demonstrated [ xxxix ].  
     [0291] Binding sites and structural requirements not fully determined, although no sequences 5′ of cleavage site are required. Folded ribozyme contains a pseudoknot structure [ xl ].  
     [0292] Reaction mechanism: attack by 2′-OH 5′ to the scissile bond to generate cleavage products with 2 ′,3′-cyclic phosphate and 5′-OH ends.  
     [0293] Only 2 known members of this class. Found in human HDV.  
     [0294] Circular form of HDV is active and shows increased nuclease stability [ xh ]  
               TABLE II                          2.5 μmol RNA Synthesis Cycle                                                 Wait           Reagent   Equivalents   Amount   Time*                                                     Phosphoramidites    6.5   163   μL   2.5           S-Ethyl Tetrazole   23.8   238   μL   2.5           Acetic Anhydride   100   233   μL    5 sec           N-Methyl Imidazole   186   233   μL    5 sec           TCA   83.2   1.73   mL   21 sec           Iodine    8.0   1.18   mL   45 sec           Acetonitrile   NA   6.67   mL   NA                                  
 
     [0295]               TABLE III                          Ribozyme Selection Characteristics                             Characteristic   Number                       HCV Genome Length   9436 kb           All Hammerhead Cleavage Sites*   1300           Conserved Region Hammerhead Cleavage    23           Sites**                                                
     [0296]               TABLE IV                          Hammerhead Ribozymes Derived from Conserved Regions of the HCV Genome                         Name   Substrate   Ribozyme Sequence                                                 5′ NCR                               HCV 5-50   CUACUGU   C   UUCACGC   GCGUGAA   CUGAUGAGGCCGUUAGGCCGAA   ACAGUAG       HCV 5-67   AAAGCGU   C   UAGCCAU   AUGGCUA   CUGAUGAGGCCGUUAGGCCGAA   ACGCUUU       HCV 5-69   AGCGUCU   A   GCCAUGG   CCAUGGC   CUGAUGAGGCCGUUAGGCCGAA   AGACGCU       HCV 5-92   UGAGUGU   C   GUGCAGC   GCUGCAC   CUGAUGAGGCCGUUAGGCCGAA   ACACUCA       HCV 5-130   GAGCCAU   A   GUGGUCU   AGACCAC   CUGAUGAGGCCGUUAGGCCGAA   AUGGCUC       HCV 5-136   UAGUGGU   C   UGCGGAA   UUCCGCA   CUGAUGAGGCCGUUAGGCCGAA   ACCACUA       HCV 5-153   GGUGAGU   A   CACCGGA   UCCGGUG   CUGAUGAGGCCGUUAGGCCGAA   ACUCACC       HCV 5-180   ACCGGGU   C   CUUUCUU   AAGAAAG   CUGAUGAGGCCGUUAGGCCGAA   ACCCGGU       HCV 5-183   GGGUCCU   U   UCUUGGA   UCCAAGA   CUGAUGAGGCCGUUAGGCCGAA   AGGACCC       HCV 5-184   GGUCCUU   U   CUUGGAU   AUCCAAG   CUGAUGAGGCCGUUAGGCCGAA   AAGGACC       HCV 5-258   GUUGGGU   C   GCGAAAG   CUUUCGC   CUGAUGAGGCCGUUAGGCCGAA   ACCCAAC       HCV 5-270   AAGGCCU   U   GUGGUAC   GUACCAC   CUGAUGAGGCCGUUAGGCCGAA   AGGCCUU       HCV 5-294   GGGUGCU   U   GCGAGUG   CACUCGC   CUGAUGAGGCCGUUAGGCCGAA   AGCACCC       HCV 5-313   GGGAGGU   C   UCGUAGA   UCUACGA   CUGAUGAGGCCGUUAGGCCGAA   ACCUCCC       HCV 5-315   GAGGUCU   C   GUAGACC   GGUCUAC   CUGAUGAGGCCGUUAGGCCGAA   AGACCUC       HCV 5-318   GUCUCGU   A   GACCGUG   CACGGUC   CUGAUGAGGCCGUUAGGCCGAA   ACGAGAC       Core Region       HCV C-30   UAAACCU   C   AAAGAAA   UUUCUUU   CUGAUGAGGCCGUUAGGCCGAA   AGGUUUA       HCV C-48   CAAACGU   A   ACACCAA   UUGGUGU   CUGAUGAGGCCGUUAGGCCGAA   ACGUUUG       HCV C-60   CAACCGU   C   GCCCACA   UGUGGGC   CUGAUGAGGCCGUUAGGCCGAA   ACGGUUG       HCV C-175   GAGCGGU   C   ACAACCU   AGGUUGU   CUGAUGAGGCCGUUAGGCCGAA   ACCGCUC       HCV C-374   GUAAGGU   C   AUCGAUA   UAUCGAU   CUGAUGAGGCCGUUAGGCCGAA   ACCUUAC       3′ NCR       HCV 3-118   UUUUUUU   U   UUUUUUU   AAAAAAA   CUGAUGAGGCCGUUAGGCCGAA   AAAAAAA       HCV 3-145   GGUGGCU   C   CAUCUUA   UAAGAUG   CUGAUGAGGCCGUUAGGCCGAA   AGCCACC       HCV 3-149   GCUCCAU   C   UUAGCCC   GGGCUAA   CUGAUGAGGCCGUUAGGCCGAA   AUGGAGC       HCV 3-151   UCCAUCU   U   AGCCCUA   UAGGGCU   CUGAUGAGGCCGUUAGGCCGAA   AGAUGGA       HCV 3-152   CCAUCUU   A   GCCCUAG   CUAGGGC   CUGAUGAGGCCGUUAGGCCGAA   AAGAUGG       HCV 3-158   UAGCCCU   A   GUCACGG   CCGUGAC   CUGAUGAGGCCGUUAGGCCGAA   AGGGCUA       HCV 3-161   CCCUAGU   C   ACGGCUA   UAGCCGU   CUGAUGAGGCCGUUAGGCCGAA   ACUAGGG       HCV 3-168   CACGGCU   A   GCUGUGA   UCACAGC   CUGAUGAGGCCGUUAGGCCGAA   AGCCGUG       HCV 3-181   GAAAGGU   C   CGUGAGC   GCUCACG   CUGAUGAGGCCGUUAGGCCGAA   ACCUUUC                    
     [0297]               TABLE V                          HCV Hammerhead Ribozyme and Target Sequence                                         Nt.               No.   Name   Pos.   Hammerhead Ribozyme   Substrate                                                                 1   HCV-27   27   UAUGGUG   CUGAUGAG   X   CGAA   AGUGUCG   CGACACU   C   CACCAUA       2   HCV-114   114   GGUCCUG   CUGAUGAG   X   CGAA   AGGCUGC   GCAGCCU   C   CAGGACC       3   HCV-128   128   CUCCCGG   CUGAUGAG   X   CGAA   AGGGGGG   CCCCCCU   C   CCGGGAG       4   HCV-148   148   UUCCGCA   CUGAUGAG   X   CGAA   ACCACUA   UAGUGGU   C   UGCGGAA       5   HCV-165   165   UCCUGUG   CUGAUGAG   X   CGAA   ACUCACC   GGUGAGU   A   CACCGGA       6   HCV-175   175   UCCUGGC   CUGAUGAG   X   CGAA   AUUCCGG   CCGGAAU   U   GCCAGGA       7   HCV-199   199   UUGAUCC   CUGAUGAG   X   CGAA   AGAAAGG   CCUUUCU   U   GGAUCAA       8   HCV-213   213   AGGCAUU   CUGAUGAG   X   CGAA   AGCGGGU   ACCCGCU   C   AAUGCCU       9   HCV-252   252   ACUCGGC   CUGAUGAG   X   CGAA   AGCAGUC   GACUGCU   A   GCCGAGU       10   HCV-260   260   CCAACAC   CUGAUGAG   X   CGAA   ACUCGGC   GCCGAGU   A   GUGUUGG       11   HCV-265   265   GCGACCC   CUGAUGAG   X   CGAA   ACACUAC   GUAGUGU   U   GGGUCGC       12   HCV-270   270   CUUUCGC   CUGAUGAG   X   CGAA   ACCCAAC   GUUGGGU   C   GCGAAAG       13   HCV-288   288   CAGGCAG   CUGAUGAG   X   CGAA   ACCACAA   UUGUGGU   A   CUGCCUG       14   HCV-298   298   AGCACCC   CUGAUGAG   X   CGAA   AUCAGGC   GCCUGAU   A   GGGUGCU       15   HCV-306   306   CACUCGC   CUGAUGAG   X   CGAA   AGCACCC   GGGUGCU   U   GCGAGUG       16   HCV-325   325   UCUACGA   CUGAUGAG   X   CGAA   ACCUCCC   GGGAGGU   C   UCGUAGA       17   HCV-327   327   GGUCUAC   CUGAUGAG   X   CGAA   AGACCUC   GAGGUCU   C   GUAGACC       18   HCV-330   330   CACGGUC   CUGAUGAG   X   CGAA   ACGAGAC   GUCUCGU   A   GACCGUG       19   HCV-407   407   GGAACUU   CUGAUGAG   X   CGAA   ACGUCCU   AGGACGU   C   AAGUUCC       20   HCV-412   412   GCCCGGG   CUGAUGAG   X   CGAA   ACUUGAC   GUCAAGU   U   CCCGGGC       21   HCV-413   413   CGCCCGG   CUGAUGAG   X   CGAA   AACUUGA   UCAAGUU   C   CCGGGCG       22   HCV-426   426   ACGAUCU   CUGAUGAG   X   CGAA   ACCACCG   CGGUGGU   C   AGAUCGU       23   HCV-472   472   CACACCC   CUGAUGAG   X   CGAA   ACGUGGG   CCCACGU   U   GGGUGUG       24   HCV-489   489   GUCUUCC   CUGAUGAG   X   CGAA   AGUCGCG   CGCGACU   A   GGAAGAC       25   HCV-498   498   CGUUCGG   CUGAUGAG   X   CGAA   AGUCUUC   GAAGACU   U   CCGAACG       26   HCV-499   499   CCGUUCG   CUGAUGAG   X   CGAA   AAGUCUU   AAGACUU   C   CGAACGG       27   HCV-508   508   AGGUUGC   CUGAUGAG   X   CGAA   ACCGUUC   GAACGGU   C   GCAACCU       28   HCV-534   534   UUGGGGA   CUGAUGAG   X   CGAA   AGGUUGU   ACAACCU   A   UCCCCAA       29   HCV-536   536   CCUUGGG   CUGAUGAG   X   CGAA   AUAGGUU   AACCUAU   C   CCCAAGG       30   HCV-546   546   GGUCGGC   CUGAUGAG   X   CGAA   AGCCUUG   CAAGGCU   C   GCCGACC       31   HCV-561   561   CAGGCCC   CUGAUGAG   X   CGAA   ACCCUCG   CGAGGGU   A   GGGCCUG       32   HCV-573   573   CCAGGCU   CUGAUGAG   X   CGAA   AGCCCAG   CUGGGCU   C   AGCCUGG       33   HCV-583   583   CCAAGGG   CUGAUGAG   X   CGAA   ACCCAGG   CCUGGGU   A   CCCUUGG       34   HCV-588   588   AGGGGCC   CUGAUGAG   X   CGAA   AGGGUAC   GUACCCU   U   GGCCCCU       35   HCV-596   596   UGCCAUA   CUGAUGAG   X   CGAA   AGGGGCC   GGCCCCU   C   UAUGGCA       36   HCV-598   598   AUUGCCA   CUGAUGAG   X   CGAA   AGAGGGG   CCCCUCU   A   UGGCAAU       37   HCV-632   632   GUGACAG   CUGAUGAG   X   CGAA   AGCCAUC   GAUGGCU   C   CUGUCAC       38   HCV-637   637   GCGGGGU   CUGAUGAG   X   CGAA   ACAGGAG   CUCCUGU   C   ACCCCGC       39   HCV-649   649   AGGCCGG   CUGAUGAG   X   CGAA   AGCCGCG   CGCGGCU   C   CCGGCCU       40   HCV-657   657   CCCCAAC   CUGAUGAG   X   CGAA   AGGCCGG   CCGGCCU   A   GUUGGGG       41   HCV-660   660   GGGCCCC   CUGAUGAG   X   CGAA   ACUAGGC   GCCUAGU   U   GGGGCCC       42   HCV-696   696   UUACCCA   CUGAUGAG   X   CGAA   AUUGCGC   GCGCAAU   C   UGGGUAA       43   HCV-707   707   UAUCGAU   CUGAUGAG   X   CGAA   ACCUUAC   GUAAGGU   C   AUCGAUA       44   HCV-710   710   GGGUAUC   CUGAUGAG   X   CGAA   AUGACCU   AGGUCAU   C   GAUACCC       45   HCV-714   714   GUGAGGG   CUGAUGAG   X   CGAA   AUCCAUG   CAUCGAU   A   CCCUCAC       46   HCV-730   730   GUCGGCG   CUGAUGAG   X   CGAA   AGCCGCA   UGCGGCU   U   CGCCGAC       47   HCV-731   731   GGUCGGC   CUGAUGAG   X   CGAA   AAGCCGC   GCGGCUU   C   GCCGACC       48   HCV-748   748   CGGAAUG   CUGAUGAG   X   CGAA   ACCCCAU   AUGGGGU   A   CAUUCCG       49   HCV-752   752   CGAGCGG   CUGAUGAG   X   CGAA   AUGUACC   GGUACAU   U   CCGCUCG       50   HCV-753   753   ACGAGCG   CUGAUGAG   X   CGAA   AAUGUAC   GUACAUU   C   CGCUCGU       51   HCV-758   758   CGCCGAC   CUGAUGAG   X   CGAA   AGCGGAA   UUCCGCU   C   GUCGGCG       52   HCV-761   761   GGGCGCC   CUGAUGAG   X   CGAA   ACGAGCG   CGCUCGU   C   GGCGCCC       53   HCV-773   773   CGCCCCC   CUGAUGAG   X   CGAA   AGGGGGG   CCCCCCU   A   GGGGGCG       54   HCV-806   806   GAACCCG   CUGAUGAG   X   CGAA   ACACCAU   AUGGUGU   C   CGGGUUC       55   HCV-812   812   CCUCCAG   CUGAUGAG   X   CGAA   ACCCGGA   UCCGGGU   U   CUGGAGG       56   HCV-813   813   UCCUCCA   CUGAUGAG   X   CGAA   AACCCGG   CCGGGUU   C   UGGAGGA       57   HCV-832   832   UGUUGCG   CUGAUGAG   X   CGAA   AGUUCAC   GUGAACU   A   CGCAACA       58   HCV-847   847   ACCGGGC   CUGAUGAG   X   CGAA   AGUUCCC   GGGAACU   U   GCCCGGU       59   HCV-855   855   AAAGAGC   CUGAUGAG   X   CGAA   ACCGGGC   GCCCGGU   U   GCUCUUU       60   HCV-859   859   AGAGAAA   CUGAUGAG   X   CGAA   AGCAACC   GGUUGCU   C   UUUCUCU       61   HCV-982   982   UGCCUCA   CUGAUGAG   X   CGAA   ACACAAU   AUUGUGU   A   UGAGGCA       62   HCV-1001   1001   UAUGCAU   CUGAUGAG   X   CGAA   AUCAUGC   GCAUGAU   C   AUGCAUA       63   HCV-1022   1022   CGCAGGG   CUGAUGAG   X   CGAA   ACGCACC   GGUGCGU   A   CCCUGCG       64   HCV-1031   1031   UCUCCCG   CUGAUGAG   X   CGAA   ACGCAGG   CCUGCGU   U   CGGGAGA       65   HCV-1032   1032   UUCUCCC   CUGAUGAG   X   CGAA   AACGCAG   CUGCGUU   C   GGGAGAA       66   HCV-1048   1048   ACAACGG   CUGAUGAG   X   CGAA   AGGCGUU   AACGCCU   C   CCGUUGU       67   HCV-1053   1053   ACCCAAC   CUGAUGAG   X   CGAA   ACGGGAG   CUCCCGU   U   GUUGGGU       68   HCV-1056   1056   GCUACCC   CUGAUGAG   X   CGAA   ACAACGG   CCGUUGU   U   GGGUAGC       69   HCV-1061   1061   UGAGCGC   CUGAUGAG   X   CGAA   ACCCAAC   GUUGGGU   A   GCGCUCA       70   HCV-1127   1127   GCAAGUC   CUGAUGAG   X   CGAA   ACGUGGC   GCCACGU   C   GACUUGC       71   HCV-1132   1132   AACGAGC   CUGAUGAG   X   CGAA   AGUCGAC   GUCGACU   U   GCUCGUU       72   HCV-1136   1136   CCCCAAC   CUGAUGAG   X   CGAA   AGCAAGU   ACUUGCU   C   GUUGGGG       73   HCV-1139   1139   CCGCCCC   CUGAUGAG   X   CGAA   ACGAGCA   UGCUCGU   U   GGGGCGG       74   HCV-1153   1153   GGAACAG   CUGAUGAG   X   CGAA   AAGCGGC   GCCGCUU   U   CUGUUCC       75   HCV-1154   1154   CGGAACA   CUGAUGAG   X   CGAA   AAAGCGG   CCGCUUU   C   UGUUCCG       76   HCV-1158   1158   AUGGCGG   CUGAUGAG   X   CGAA   ACAGAAA   UUUCUGU   U   CCGCCAU       77   HCV-1159   1159   CAUGGCG   CUGAUGAG   X   CGAA   AACAGAA   UUCUGUU   C   CGCCAUG       78   HCV-1168   1168   CCCCACG   CUGAUGAG   X   CGAA   ACAUGGC   GCCAUGU   A   CGUGGGG       79   HCV-1189   1189   GAAAACG   CUGAUGAG   X   CGAA   AUCCGCA   UGCGGAU   C   CGUUUUC       80   HCV-1193   1193   CGAGGAA   CUGAUGAG   X   CGAA   ACGGAUC   GAUCCGU   U   UUCCUCG       81   HCV-1194   1194   ACGAGGA   CUGAUGAG   X   CGAA   AACGGAU   AUCCGUU   U   UCCUCGU       82   HCV-1195   1195   GACGAGG   CUGAUGAG   X   CGAA   AAACGGA   UCCGUUU   U   CCUCGUC       83   HCV-1196   1196   AGACGAG   CUGAUGAG   X   CGAA   AAAACGG   CCGUUUU   C   CUCGUCU       84   HCV-1280   1280   GACCUGA   CUGAUGAG   X   CGAA   ACAUGGC   GCCAUGU   A   UCAGGUC       85   HCV-1282   1282   GUGACCU   CUGAUGAG   X   CGAA   AUACAUG   CAUGUAU   C   AGGUCAC       86   HCV-1287   1287   AUGCGGU   CUGAUGAG   X   CGAA   ACCUGAU   AUCAGGU   C   ACCGCAU       87   HCV-1373   1373   UAUCCAC   CUGAUGAG   X   CGAA   ACAGCUU   AAGCUGU   C   GUGGAUA       88   HCV-1380   1380   GCCACCA   CUGAUGAG   X   CGAA   AUCCACG   CGUGGAU   A   UGGUGGC       89   HCV-1406   1406   CCGCUAG   CUGAUGAG   X   CGAA   ACUCCCC   GGGGAGU   C   CUAGCGG       90   HCV-1409   1409   GGCCCGC   CUGAUGAG   X   CGAA   AGGACUC   GAGUCCU   A   GCGGGCC       91   HCV-1418   1418   AGUAGGC   CUGAUGAG   X   CGAA   AGGCCCG   CGGGCCU   U   GCCUACU       92   HCV-1423   1423   GGAAUAG   CUGAUGAG   X   CGAA   AGGCAAG   CUUGCCU   A   CUAUUCC       93   HCV-1426   1426   CAUGGAA   CUGAUGAG   X   CGAA   AGUAGGC   GCCUACU   A   UUCCAUG       94   HCV-1428   1428   ACCAUGG   CUGAUGAG   X   CGAA   AUAGUAG   CUACUAU   U   CCAUGGU       95   HCV-1429   1429   CACCAUG   CUGAUGAG   X   CGAA   AAUAGUA   UACUAUU   C   CAUGGUG       96   HCV-1727   1727   ACUUGUC   CUGAUGAG   X   CGAA   AUGGAGC   GCUCCAU   C   GACAAGU       97   HCV-1735   1735   CUGAGCG   CUGAUGAG   X   CGAA   ACUUGUC   GACAAGU   U   CGCUCAG       98   HCV-1736   1736   CCUGAGC   CUGAUGAG   X   CGAA   AACUUGU   ACAAGUU   C   GCUCAGG       99   HCV-1740   1740   CAUCCCU   CUGAUGAG   X   CGAA   AGCGAAC   GUUCGCU   C   AGGGAUG       100   HCV-1757   1757   UAUAGGU   CUGAUGAG   X   CGAA   AUGGGGC   GCCCCAU   C   ACCUAUA       101   HCV-1762   1762   CUCGGUA   CUGAUGAG   X   CGAA   AGGUGAU   AUCACCU   A   UACCGAG       102   HCV-1795   1795   CCAGCAG   CUGAUGAG   X   CGAA   AAGGCCU   AGGCCUU   A   CUGCUGG       103   HCV-1806   1806   GGUGCGU   CUGAUGAG   X   CGAA   AUGCCAG   CUGGCAU   U   ACGCACC       104   HCV-1807   1807   AGGUGCG   CUGAUGAG   X   CGAA   AAUGCCA   UGGCAUU   A   CGCACCU       105   HCV-1815   1815   CACUGCC   CUGAUGAG   X   CGAA   AGGUGCG   CGCACCU   C   GGCAGUG       106   HCV-1827   1827   GGUACGA   CUGAUGAG   X   CGAA   ACCACAC   GUGUGGU   A   UCGUACC       107   HCV-1829   1829   CAGGUAC   CUGAUGAG   X   CGAA   AUACCAC   GUGGUAU   C   GUACCUG       108   HCV-1832   1832   ACGCAGG   CUGAUGAG   X   CGAA   ACGAUAC   GUAUCGU   A   CCUGCGU       109   HCV-1840   1840   CACCUGC   CUGAUGAG   X   CGAA   ACGCAGG   CCUGCGU   C   GCAGGUG       110   HCV-1854   1854   UACACUG   CUGAUGAG   X   CGAA   ACCACAC   GUGUGGU   C   CAGUGUA       111   HCV-1883   1883   CCACUAC   CUGAUGAG   X   CGAA   ACAGGGC   GCCCUGU   U   GUAGUGG       112   HCV-1886   1886   UCCCCAC   CUGAUGAG   X   CGAA   ACAACAG   CUGUUGU   A   GUGGGGA       113   HCV-1902   1902   CCGGACC   CUGAUGAG   X   CGAA   AUCGGUC   GACCGAU   C   GGUCCGG       114   HCV-1906   1906   GGCACCG   CUGAUGAG   X   CGAA   ACCGAUC   GAUCGGU   C   CGGUGCC       115   HCV-1917   1917   UUAUACG   CUGAUGAG   X   CGAA   AGGGGCA   UGCCCCU   A   CGUAUAA       116   HCV-1921   1921   CCAGUUA   CUGAUGAG   X   CGAA   ACGUAGG   CCUACGU   A   UAACUGG       117   HCV-1923   1923   CCCCAGU   CUGAUGAG   X   CGAA   AUACGUA   UACGUAU   A   ACUGGGG       118   HCV-1990   1990   ACAGCCA   CUGAUGAG   X   CGAA   ACCAGUU   AACUGGU   U   UGGCUGU       119   HCV-1991   1991   UACAGCC   CUGAUGAG   X   CGAA   AACCAGU   ACUGGUU   U   GGCUGUA       120   HCV-1998   1998   AUCCAUG   CUGAUGAG   X   CGAA   ACAGCCA   UGGCUGU   A   CAUGGAU       121   HCV-2043   2043   UUGCACG   CUGAUGAG   X   CGAA   AGGGCCC   GGGCCCU   C   CGUGCAA       122   HCV-2054   2054   CCCCCCC   CUGAUGAG   X   CGAA   AUGUUGC   GCAACAU   C   GGGGGGG       123   HCV-2063   2083   GGUUGCC   CUGAUGAG   X   CGAA   ACCCCCC   GGGGGGU   C   GGCAACC       124   HCV-2072   2072   UCAAGGU   CUGAUGAG   X   CGAA   AGGUUGC   GCAACCU   C   ACCUUGA       125   HCV-2077   2077   GCAGGUC   CUGAUGAG   X   CGAA   AGGUGAG   CUCACCU   U   GACCUGC       126   HCV-2121   2121   UUUGUGU   CUGAUGAG   X   CGAA   AGUGGCC   GGCCACU   U   ACACAAA       127   HCV-2122   2122   UUUUGUG   CUGAUGAG   X   CGAA   AAGUGGC   GCCACUU   A   CACAAAA       128   HCV-2137   2137   UGGCCCC   CUGAUGAG   X   CGAA   AGCCACA   UGUGGCU   C   GGGGCCA       129   HCV-2149   2149   AGGUGUU   CUGAUGAG   X   CGAA   ACCAUGG   CCAUGGU   U   AACACCU       130   HCV-2150   2150   UAGGUGU   CUGAUGAG   X   CGAA   AACCAUG   CAUGGUU   A   ACACCUA       131   HCV-2219   2219   CCUUAAA   CUGAUGAG   X   CGAA   AUGGUAA   UUACCAU   C   UUUAAGG       132   HCV-2221   2221   AACCUUA   CUGAUGAG   X   CGAA   AGAUGGU   ACCAUCU   U   UAAGGUU       133   HCV-2261   2261   CAGCACU   CUGAUGAG   X   CGAA   AGCCUGU   ACAGGCU   U   AGUGCUG       134   HCV-2262   2262   GCAGCAC   CUGAUGAG   X   CGAA   AAGCCUG   CAGGCUU   A   GUGCUGC       135   HCV-2295   2295   AGGUCGC   CUGAUGAG   X   CGAA   ACGCUCU   AGAGCGU   U   GCGACCU       136   HCV-2320   2320   GAGCUCC   CUGAUGAG   X   CGAA   AUCUGUC   GACAGAU   C   GGAGCUC       137   HCV-2327   2327   GCGGGCU   CUGAUGAG   X   CGAA   AGCUCCG   CGGAGCU   C   AGCCCGC       138   HCV-2344   2344   UGUCGUG   CUGAUGAG   X   CGAA   ACAGCAG   CUGCUGU   C   CACGACA       139   HCV-2417   2417   UCUGAUG   CUGAUGAG   X   CGAA   AGGUGGA   UCCACCU   C   CAUCAGA       140   HCV-2421   2421   AUGUUCU   CUGAUGAG   X   CGAA   AUGGAGG   CCUCCAU   C   AGAACAU       141   HCV-2429   2429   CGUCCAC   CUGAUGAG   X   CGAA   AUGUUCU   AGAACAU   C   GUGGACG       142   HCV-2534   2534   AGGCACA   CUGAUGAG   X   CGAA   ACGCGCG   CGCGCGU   C   UGUGCCU       143   HCV-2585   2585   GGUUCUC   CUGAUGAG   X   CGAA   AGGGCGG   CCGCCCU   A   GAGAACC       144   HCV-2600   2600   CGUUGAG   CUGAUGAG   X   CGAA   ACCACCA   UGGUGGU   C   CUCAACG       145   HCV-2603   2603   CCGCGUU   CUGAUGAG   X   CGAA   AGGACCA   UGGUCCU   C   AACGCGG       146   HCV-2671   2671   CUUGAUG   CUGAUGAG   X   CGAA   ACCAGGC   GCCUGGU   A   CAUCAAG       147   HCV-2675   2675   UGCCCUU   CUGAUGAG   X   CGAA   AUGUACC   GGUACAU   C   AAGGGCA       148   HCV-2690   2690   CCCCAGG   CUGAUGAG   X   CGAA   ACCAGCC   GGCUGGU   C   CCUGGGG       149   HCV-2704   2704   CAGAGCA   CUGAUGAG   X   CGAA   AUGCCGC   GCGGCAU   A   UGCUCUG       150   HCV-2709   2709   CCGUACA   CUGAUGAG   X   CGAA   AGCAUAU   AUAUGCU   C   UGUACGG       151   HCV-2713   2713   CACGCCG   CUGAUGAG   X   CGAA   ACAGAGC   GCUCUGU   A   CGGCGUG       152   HCV-2738   2738   CCAGCAG   CUGAUGAG   X   CGAA   ACCAGGA   UCCUGCU   C   CUGCUGG       153   HCV-2763   2763   AUGGCGU   CUGAUGAG   X   CGAA   AGCCCGU   ACGGGCU   U   ACGCCAU       154   HCV-2764   2764   CAUGGCG   CUGAUGAG   X   CGAA   AAGCCCG   CGGGCUU   A   CGCCAUG       155   HCV-2878   2878   GUAUUGU   CUGAUGAG   X   CGAA   ACCACCA   UGGUGGU   U   ACAAUAC       156   HCV-2879   2879   AGUAUUG   CUGAUGAG   X   CGAA   AACCACC   GGUGGUU   A   CAAUACU       157   HCV-2884   2884   GAUAAAG   CUGAUGAG   X   CGAA   AUUGUAA   UUACAAU   A   CUUUAUC       158   HCV-2887   2887   GGUGAUA   CUGAUGAG   X   CGAA   AGUAUUG   CAAUACU   U   UAUCACC       159   HCV-2888   2888   UGGUGAU   CUGAUGAG   X   CGAA   AAGUAUU   AAUACUU   U   AUCACCA       160   HCV-2910   2910   ACGCACA   CUGAUGAG   X   CGAA   AUGCGCC   GGCGCAU   U   UGUGCGU       161   HCV-2911   2911   CACGCAC   CUGAUGAG   X   CGAA   AAUGCGC   GCGCAUU   U   GUGCGUG       162   HCV-2924   2924   GAGGGGG   CUGAUGAG   X   CGAA   ACCCACA   UGUGGGU   C   CCCCCUC       163   HCV-2931   2931   ACAUUGA   CUGAUGAG   X   CGAA   AGGGGGG   CCCCCCU   C   UCAAUGU       164   HCV-2933   2933   GGACAUU   CUGAUGAG   X   CGAA   AGAGGGG   CCCCUCU   C   AAUGUCC       165   HCV-2939   2939   CCCCCCG   CUGAUGAG   X   CGAA   ACAUUGA   UCAAUGU   C   CGGGGGG       166   HCV-2958   2958   AGGAUGA   CUGAUGAG   X   CGAA   AGCAUCG   CGAUGCU   A   UCAUCCU       167   HCV-2960   2960   GGAGGAU   CUGAUGAG   X   CGAA   AUAGCAU   AUGCUAU   C   AUCCUCC       168   HCV-2963   2963   UGAGGAG   CUGAUGAG   X   CGAA   AUGAUAG   CUAUCAU   C   CUCCUCA       169   HCV-2966   2966   AUGUGAG   CUGAUGAG   X   CGAA   AGGAUGA   UCAUCCU   C   CUCACAU       170   HCV-2969   2969   CACAUGU   CUGAUGAG   X   CGAA   AGGAGGA   UCCUCCU   C   ACAUGUG       171   HCV-3059   3059   UCGCAGU   CUGAUGAG   X   CGAA   AUGGCAG   CUGCCAU   A   ACUGCGA       172   HCV-3138   3138   UGGACGU   CUGAUGAG   X   CGAA   AUGGCCU   AGGCCAU   U   ACGUCCA       173   HCV-3139   3139   UUGGACG   CUGAUGAG   X   CGAA   AAUGGCC   GGCCAUU   A   CGUCCAA       174   HCV-3143   3143   CCAUUUG   CUGAUGAG   X   CGAA   ACGUAAU   AUUACGU   C   CAAAUGG       175   HCV-3154   3154   CUUCAUG   CUGAUGAG   X   CGAA   AGGCCAU   AUGGCCU   U   CAUGAAG       176   HCV-3155   3155   GCUUCAU   CUGAUGAG   X   CGAA   AAGGCCA   UGGCCUU   C   AUGAAGC       177   HCV-3209   3209   AAUCCUG   CUGAUGAG   X   CGAA   AGCGGGG   CCCCGCU   A   CAGGAUU       178   HCV-3216   3216   UGGGCCC   CUGAUGAG   X   CGAA   AUCCUGU   ACAGGAU   U   GGGCCCA       179   HCV-3233   3233   GGUCUCG   CUGAUGAG   X   CGAA   AGGCCCG   CGGGCCU   A   CGAGACC       180   HCV-3242   3242   CCACCGC   CUGAUGAG   X   CGAA   AGGUCUC   GAGACCU   U   GCGGUGG       181   HCV-3263   3263   AGAAGAC   CUGAUGAG   X   CGAA   ACGGGCU   AGCCCGU   C   GUCUUCU       182   HCV-3266   3266   CAGAGAA   CUGAUGAG   X   CGAA   ACGACGG   CCGUCGU   C   UUCUCUG       183   HCV-3268   3268   GUCAGAG   CUGAUGAG   X   CGAA   AGACGAC   GUCGUCU   U   CUCUGAC       184   HCV-3290   3290   AGGUGAU   CUGAUGAG   X   CGAA   AUCUUGG   CCAAGAU   C   AUCACCU       185   HCV-3293   3293   CCCAGGU   CUGAUGAG   X   CGAA   AUGAUCU   AGAUCAU   C   ACCUGGG       186   HCV-3329   3329   CCAAGAU   CUGAUGAG   X   CGAA   AUGUCCC   GGGACAU   C   AUCUUGG       187   HCV-3332   3332   GUCCCAA   CUGAUGAG   X   CGAA   AUGAUGU   ACAUCAU   C   UUGGGAC       188   HCV-3334   3334   CAGUCCC   CUGAUGAG   X   CGAA   AGAUGAU   AUCAUCU   U   GGGACUG       189   HCV-3347   3347   GGGCGGA   CUGAUGAG   X   CGAA   ACGGGCA   UGCCCGU   C   UCCGCCC       190   HCV-3349   3349   UCGGGCG   CUGAUGAG   X   CGAA   ACACGGG   CCCGUCU   C   CGCCCGA       191   HCV-3371   3371   CCAGAAG   CUGAUGAG   X   CGAA   AUCUCCC   GGGAGAU   A   CUUCUGG       192   HCV-3416   3416   GGGCAAG   CUGAUGAG   X   CGAA   AGUCGCC   GGCGACU   C   CUUGCCC       193   HCV-3419   3419   UGGGGGC   CUGAUGAG   X   CGAA   AGGAGUC   GACUCCU   U   GCCCCCA       194   HCV-3428   3428   AGGCCGU   CUGAUGAG   X   CGAA   AUGGGGG   CCCCCAU   C   ACGGCCU       195   HCV-3482   3482   GGCCUGU   CUGAUGAG   X   CGAA   AGUCUAG   CUAGCCU   C   ACAGGCC       196   HCV-3518   3518   CCACUUG   CUGAUGAG   X   CGAA   ACCUCCC   GGGAGGU   U   CAAGUGG       197   HCV-3519   3519   ACCACUU   CUGAUGAG   X   CGAA   AACCUCC   GGAGGUU   C   AAGUGGU       198   HCV-3527   3527   CGGUGGA   CUGAUGAG   X   CGAA   ACCACUU   AAGUGGU   U   UCCACCG       199   HCV-3528   3528   GCGGUGG   CUGAUGAG   X   CGAA   AACCACU   AGUGGUU   U   CCACCGC       200   HCV-3529   3529   UGCGGUG   CUGAUGAG   X   CGAA   AAACCAC   GUGGUUU   C   CACCGCA       201   HCV-3576   3576   ACGGUCC   CUGAUGAG   X   CGAA   ACACACA   UGUGUGU   U   GGACCGU       202   HCV-3601   3601   GGUCUUU   CUGAUGAG   X   CGAA   AGCCGGC   GCCGGCU   C   AAAGACC       203   HCV-3611   3611   GGCCGGC   CUGAUGAG   X   CGAA   AGGGUCU   AGACCCU   A   GCCGGCC       204   HCV-3684   3684   GCCCCGG   CUGAUGAG   X   CGAA   AGGCGCA   UGCGCCU   C   CCGGGGC       205   HCV-3696   3696   GUAAGGG   CUGAUGAG   X   CGAA   ACGCGCC   GGCGCGU   U   CCCUUAC       206   HCV-3697   3697   UGUAAGG   CUGAUGAG   X   CGAA   AACGCGC   GCGCGUU   C   CCUUACA       207   HCV-3701   3701   AUGGUGU   CUGAUGAG   X   CGAA   AGGGAAC   GUUCCCU   U   ACACCAU       208   HCV-3702   3702   CAUGGUG   CUGAUGAG   X   CGAA   AAGGGAA   UUCCCUU   A   CACCAUG       209   HCV-3724   3724   GAGGUCC   CUGAUGAG   X   CGAA   AGCUACC   GGUAGCU   C   GGACCUC       210   HCV-3731   3731   CCAGAUA   CUGAUGAG   X   CGAA   AGGUCCG   CGGACCU   C   UAUCUGG       211   HCV-3733   3733   GACCAGA   CUGAUGAG   X   CGAA   AGAGGUC   GACCUCU   A   UCUGGUC       212   HCV-3735   3735   GUGACCA   CUGAUGAG   X   CGAA   AUAGAGG   CCUCUAU   C   UGGUCAC       213   HCV-3740   3740   GUCUCGU   CUGAUGAG   X   CGAA   ACCAGAU   AUCUGGU   C   ACGAGAC       214   HCV-3761   3761   GCACCGG   CUGAUGAG   X   CGAA   AUGACGU   ACGUCAU   U   CCGGUGC       215   HCV-3762   3762   CGCACCG   CUGAUGAG   X   CGAA   AAUGACG   CGUCAUU   C   CGGUGCG       216   HCV-3786   3786   CUCCCCC   CUGAUGAG   X   CGAA   ACCGUCA   UGACGGU   C   GGGGGAG       217   HCV-3797   3797   GGGACAG   CUGAUGAG   X   CGAA   AGGCUCC   GGAGCCU   A   CUGUCCC       218   HCV-3802   3802   UCUGGGG   CUGAUGAG   X   CGAA   ACAGUAG   CUACUGU   C   CCCCAGA       219   HCV-3835   3835   GCCACCC   CUGAUGAG   X   CGAA   AAGAGCC   GGCUCUU   C   GGGUGGC       220   HCV-3851   3851   AAGGGCA   CUGAUGAG   X   CGAA   AGCAGUG   CACUGCU   C   UGCCCUU       221   HCV-3858   3858   UGCCCCG   CUGAUGAG   X   CGAA   AGGGCAG   CUGCCCU   U   CGGGGCA       222   HCV-3859   3859   GUGCCCC   CUGAUGAG   X   CGAA   AAGGGCA   UGCCCUU   C   GGGGCAC       223   HCV-3872   3872   AGAUGCC   CUGAUGAG   X   CGAA   ACAGCGU   ACGCUGU   A   GGCAUCU       224   HCV-3878   3878   CCCGGAA   CUGAUGAG   X   CGAA   AUGCCUA   UAGGCAU   C   UUCCGGG       225   HCV-3880   3880   AGCCCGG   CUGAUGAG   X   CGAA   AGAUGCC   GGCAUCU   U   CCGGGCU       226   HCV-3881   3881   CAGCCCG   CUGAUGAG   X   CGAA   AAGAUGC   GCAUCUU   C   CGGGCUG       227   HCV-3908   3908   CCUUCGC   CUGAUGAG   X   CGAA   ACCCCCC   GGGGGGU   U   GCGAAGG       228   HCV-4056   4056   GGCACUU   CUGAUGAG   X   CGAA   AGUGCUC   GAGCACU   A   AAGUGCC       229   HCV-4072   4072   GGCUGCG   CUGAUGAG   X   CGAA   ACGCAGC   GCUGCGU   A   CGCAGCC       230   HCV-4087   4087   UACCUUG   CUGAUGAG   X   CGAA   ACCCUUG   CAAGGGU   A   CAAGGUA       231   HCV-4115   4115   UGGCGGC   CUGAUGAG   X   CGAA   ACAGAUG   CAUCUGU   U   GCCGCCA       232   HCV-4175   4175   CAGUUCU   CUGAUGAG   X   CGAA   AUGUUGG   CCAACAU   C   AGAACUG       233   HCV-4187   4187   UGGUCCU   CUGAUGAG   X   CGAA   ACCCCAG   CUGGGGU   A   AGGACCA       234   HCV-4228   4228   CUUACCA   CUGAUGAG   X   CGAA   AGGUGGA   UCCACCU   A   UGGUAAG       235   HCV-4233   4233   AGGAACU   CUGAUGAG   X   CGAA   ACCAUAG   CUAUGGU   A   AGUUCCU       236   HCV-4237   4237   GGCAAGG   CUGAUGAG   X   CGAA   ACUUACC   GGUAAGU   U   CCUUGCC       237   HCV-4238   4238   CGGCAAG   CUGAUGAG   X   CGAA   AACUUAC   GUAAGUU   C   CUUGCCG       238   HCV-4241   4241   CGUCGGC   CUGAUGAG   X   CGAA   AGGAACU   AGUUCCU   U   GCCGACG       239   HCV-4280   4280   CACAUAU   CUGAUGAG   X   CGAA   AUGAUAU   AUAUCAU   A   AUAUGUG       240   HCV-4283   4283   CAUCACA   CUGAUGAG   X   CGAA   AUUAUGA   UCAUAAU   A   UGUGAUG       241   HCV-4337   4337   GGUCCAG   CUGAUGAG   X   CGAA   ACUGUGC   GCACAGU   C   CUGGACC       242   HCV-4370   4370   GCACGAC   CUGAUGAG   X   CGAA   AGCCGCG   CGCGGCU   C   GUCGUGC       243   HCV-4373   4373   CGAGCAC   CUGAUGAG   X   CGAA   ACGAGCC   GGCUCGU   C   GUGCUCG       244   HCV-4379   4379   CGGUGGC   CUGAUGAG   X   CGAA   AGCACGA   UCGUGCU   C   GCCACCG       245   HCV-4425   4425   UCCUCAA   CUGAUGAG   X   CGAA   AUUUGGG   CCCAAAU   A   UUGAGGA       246   HCV-4444   4444   AGUGUUG   CUGAUGAG   X   CGAA   ACAGAGC   GCUCUGU   C   CAACACU       247   HCV-4460   4460   AGAAGGG   CUGAUGAG   X   CGAA   AUCUCUC   GAGAGAU   C   CCCUUCU       248   HCV-4481   4481   CGAGGGG   CUGAUGAG   X   CGAA   AUGGCCU   AGGCCAU   C   CCCCUCG       249   HCV-4487   4487   UGGCCUC   CUGAUGAG   X   CGAA   AGGGGGA   UCCCCCU   C   GAGGCCA       250   HCV-4496   4496   CCCCCUU   CUGAUGAG   X   CGAA   AUGGCCU   AGGCCAU   C   AAGGGGG       251   HCV-4528   4528   CUUCUUG   CUGAUGAG   X   CGAA   AGUGGCA   UGCCACU   C   CAAGAAG       252   HCV-4577   4577   CGGCAUU   CUGAUGAG   X   CGAA   AUUCCGA   UCGGAAU   C   AAUGCCG       253   HCV-4586   4586   AAUACGC   CUGAUGAG   X   CGAA   ACGGCAU   AUGCCGU   A   GCGUAUU       254   HCV-4591   4591   CCGGUAA   CUGAUGAG   X   CGAA   ACGCUAC   GUAGCGU   A   UUACCGG       255   HCV-4593   4593   CCCCGGU   CUGAUGAG   X   CGAA   AUACGCU   AGCGUAU   U   ACCGGGG       256   HCV-4594   4594   ACCCCGG   CUGAUGAG   X   CGAA   AAUACGC   GCGUAUU   A   CCGGGGU       257   HCV-4616   4616   UCGGUAU   CUGAUGAG   X   CGAA   ACGGACA   UGUCCGU   C   AUACCGA       258   HCV-4619   4619   UAGUCGG   CUGAUGAG   X   CGAA   AUGACGG   CCGUCAU   A   CCGACUA       259   HCV-4626   4626   UCUCCGC   CUGAUGAG   X   CGAA   AGUCGGU   ACCGACU   A   GCGGAGA       260   HCV-4672   4672   ACCGGUG   CUGAUGAG   X   CGAA   AGCCCGU   ACGGGCU   A   CACCGGU       261   HCV-4697   4697   UGCAGUC   CUGAUGAG   X   CGAA   AUCACCG   CGGUGAU   C   GACUGCA       262   HCV-4789   4789   UGAGCGC   CUGAUGAG   X   CGAA   ACACCGC   GCGGUGU   C   GCGCUCA       263   HCV-4795   4795   CCGUUGU   CUGAUGAG   X   CGAA   AGCGCGA   UCGCGCU   C   ACAACGG       264   HCV-4920   4920   UCAUACC   CUGAUGAG   X   CGAA   AGCACAG   CUGUGCU   U   GGUAUGA       265   HCV-4924   4924   GAGCUCA   CUGAUGAG   X   CGAA   ACCAAGC   GCUUGGU   A   UGAGCUC       266   HCV-4931   4931   CGGGCGU   CUGAUGAG   X   CGAA   AGCUCAU   AUGAGCU   C   ACGCCCG       267   HCV-4947   4947   CUGACUG   CUGAUGAG   X   CGAA   AGUCUCA   UGAGACU   A   CAGUCAG       268   HCV-4952   4952   GCAACCU   CUGAUGAG   X   CGAA   ACUGUAG   CUACAGU   C   AGGUUGC       269   HCV-4957   4957   AGCCCGC   CUGAUGAG   X   CGAA   ACCUGAC   GUCAGGU   U   GCGGGCU       270   HCV-4965   4965   UUCAGGU   CUGAUGAG   X   CGAA   AGCCCGC   GCGGGCU   U   ACCUGAA       271   HCV-4966   4966   AUUCAGG   CUGAUGAG   X   CGAA   AAGCCCG   CGGGCUU   A   CCUGAAU       272   HCV-4974   4974   CCUGGUG   CUGAUGAG   X   CGAA   AUUCAGG   CCUGAAU   A   CACCAGG       273   HCV-4984   4984   GACGGGC   CUGAUGAG   X   CGAA   ACCCUGG   CCAGGGU   U   GCCCGUC       274   HCV-4991   4991   CCUGGCA   CUGAUGAG   X   CGAA   ACGGGCA   UGCCCGU   C   UGCCAGG       275   HCV-5004   5004   AACUCCA   CUGAUGAG   X   CGAA   AUGGUCC   GGACCAU   C   UGGAGUU       276   HCV-5102   5102   GGUAUGC   CUGAUGAG   X   CGAA   ACCAGGU   ACCUGGU   A   GCAUACC       277   HCV-5107   5107   GGCUUGG   CUGAUGAG   X   CGAA   AUGCUAC   GUAGCAU   A   CCAAGCC       278   HCV-5133   5133   GGAGCCU   CUGAUGAG   X   CGAA   AGCCCUG   CAGGGCU   C   AGGCUCC       279   HCV-5218   5218   UAGCCUA   CUGAUGAG   X   CGAA   ACAGCAG   CUGCUGU   A   UAGGCUA       280   HCV-5220   5220   CCUAGCC   CUGAUGAG   X   CGAA   AUACAGC   GCUGUAU   A   GGCUAGG       281   HCV-5306   5306   UAGUGAC   CUGAUGAG   X   CGAA   ACCUCCA   UGGAGGU   C   GUCACUA       282   HCV-5309   5309   UGCUAGU   CUGAUGAG   X   CGAA   ACGACCU   AGGUCGU   C   ACUACCA       283   HCV-5313   5313   CAGGUGC   CUGAUGAG   X   CGAA   AGUGACG   CGUCACU   A   GCACCUG       284   HCV-5330   5330   CUCCGCC   CUGAUGAG   X   CGAA   ACCAGCA   UGCUGGU   A   GGCGGAG       285   HCV-5339   5339   CUGCAAG   CUGAUGAG   X   CGAA   ACUCCGC   GCGGAGU   C   CUUGCAG       286   HCV-5342   5342   GAGCUGC   CUGAUGAG   X   CGAA   AGGACUC   GAGUCCU   U   GCAGCUC       287   HCV-5359   5359   CAGGCAA   CUGAUGAG   X   CGAA   AUGCGGC   GCCGCAU   A   UUGCCUG       288   HCV-5361   5361   GUCAGGC   CUGAUGAG   X   CGAA   AUAUGCG   CGCAUAU   U   GCCUGAC       289   HCV-5376   5376   ACCACAC   CUGAUGAG   X   CGAA   ACCGGUU   AACCGGU   A   GUGUGGU       290   HCV-5399   5399   ACAAAAU   CUGAUGAG   X   CGAA   AUCCUAC   GUAGGAU   C   AUUUUGU       291   HCV-5423   5423   CGGGAAC   CUGAUGAG   X   CGAA   ACAGCCG   CGGCUGU   U   GUUCCCG       292   HCV-5426   5426   UGUCGGG   CUGAUGAG   X   CGAA   ACAACAG   CUGUUGU   U   CCCGACA       293   HCV-5427   5427   CUGUCGG   CUGAUGAG   X   CGAA   AACAACA   UGUUGUU   C   CCGACAG       294   HCV-5524   5524   CUGCUUG   CUGAUGAG   X   CGAA   ACUGCUC   GAGCAGU   U   CAAGCAG       295   HCV-5525   5525   UCUGCUU   CUGAUGAG   X   CGAA   AACUGCU   AGCAGUU   C   AAGCAGA       296   HCV-5583   5583   ACCACGG   CUGAUGAG   X   CGAA   AGCAGCG   CGCUGCU   C   CCGUGGU       297   HCV-5596   5596   CCACCUG   CUGAUGAG   X   CGAA   ACUCCAC   GUGGAGU   C   CAGGUGG       298   HCV-5612   5612   AGGCCUC   CUGAUGAG   X   CGAA   AGGGCCC   GGGCCCU   U   GAGGCCU       299   HCV-5620   5620   UGCCCAG   CUGAUGAG   X   CGAA   AGGCCUC   GAGGCCU   U   CUGGGCA       300   HCV-5621   5621   UUGCCCA   CUGAUGAG   X   CGAA   AAGGCCU   AGGCCUU   C   UGGGCAA       301   HCV-5674   5674   AGUGGAU   CUGAUGAG   X   CGAA   AGCCUGC   GCAGGCU   U   AUCCACU       302   HCV-5675   5675   GAGUGGA   CUGAUGAG   X   CGAA   AAGCCUG   CAGGCUU   A   UCCACUC       303   HCV-5767   5767   GAUGUUG   CUGAUGAG   X   CGAA   ACAGGAG   CUCCUGU   U   CAACAUC       304   HCV-5768   5768   AGAUGUU   CUGAUGAG   X   CGAA   AACAGGA   UCCUGUU   C   AACAUCU       305   HCV-5801   5801   GAGGAGC   CUGAUGAG   X   CGAA   AGUUGAG   CUCAACU   C   GCUCCUC       306   HCV-5805   5805   CUGGGAG   CUGAUGAG   X   CGAA   AGCGAGU   ACUCGCU   C   CUCCCAG       307   HCV-5821   5821   GAAGGCC   CUGAUGAG   X   CGAA   AAGCAGC   GCUGCUU   C   GGCCUUC       308   HCV-5827   5827   GCCCACG   CUGAUGAG   X   CGAA   AGGCCGA   UCGGCCU   U   CGUGGGC       309   HCV-5828   5828   CGCCCAC   CUGAUGAG   X   CGAA   AAGGCCG   CGGCCUU   C   GUGGGCG       310   HCV-5843   5843   CACCGGC   CUGAUGAG   X   CGAA   AUGCCGG   CCGGCAU   U   GCCGGUG       311   HCV-5858   5858   UGCUGCC   CUGAUGAG   X   CGAA   AUGGCCG   CGGCCAU   U   GGCAGCA       312   HCV-5867   5867   CAAGGCC   CUGAUGAG   X   CGAA   AUCCUCC   GCAGCAU   A   GGCCUUG       313   HCV-5873   5873   CCUUCCC   CUGAUGAG   X   CGAA   AGGCCUA   UAGGCCU   U   GGGAAGG       314   HCV-5905   5905   CGCUCCA   CUGAUGAG   X   CGAA   AGCCCGC   GCGGGCU   A   UGGAGCG       315   HCV-5930   5930   AAGCCAC   CUGAUGAG   X   CGAA   AGUGCAC   GUGCACU   C   GUGGCUU       316   HCV-5937   5937   ACCUUAA   CUGAUGAG   X   CGAA   AGCCACG   CGUGGCU   U   UUAAGGU       317   HCV-5938   5938   GACCUUA   CUGAUGAG   X   CGAA   AAGCCAC   GUGGCUU   U   UAAGGUC       318   HCV-5939   5939   UGACCUU   CUGAUGAG   X   CGAA   AAAGCCA   UGGCUUU   U   AAGGUCA       319   HCV-5940   5940   AUGACCU   CUGAUGAG   X   CGAA   AAAAGCC   GGCUUUU   A   AGGUCAU       320   HCV-5945   5945   CGCUCAU   CUGAUGAG   X   CGAA   ACCUUAA   UUAAGGU   C   AUGAGCG       321   HCV-5965   5965   CUCGGCG   CUGAUGAG   X   CGAA   AGGGCGC   GCGCCCU   C   CGCCGAG       322   HCV-5981   5981   GCAAGUU   CUGAUGAG   X   CGAA   ACCAGGU   ACCUGGU   U   AACUUGC       323   HCV-5982   5982   AGCAAGU   CUGAUGAG   X   CGAA   AACCAGG   CCUGGUU   A   ACUUGCU       324   HCV-5990   5990   UGGCAGG   CUGAUGAG   X   CGAA   AGCAAGU   ACUUGCU   C   CCUGCCA       325   HCV-6004   6004   GCCGGGG   CUGAUGAG   X   CGAA   AGAGGAU   AUCCUCU   C   CCCCGGC       326   HCV-6020   6020   CCCCGAC   CUGAUGAG   X   CGAA   ACCAGGG   CCCUGGU   C   GUCGGGG       327   HCV-6023   6023   CGACCCC   CUGAUGAG   X   CGAA   ACGACCA   UGGUCGU   C   GGGGUCG       328   HCV-6029   6029   CACACAC   CUGAUGAG   X   CGAA   ACCCCGA   UCGGGGU   C   GUGUGUG       329   HCV-6044   6044   GACGCAG   CUGAUGAG   X   CGAA   AUUGCUG   CAGCAAU   C   CUGCGUC       330   HCV-6051   6051   ACGUGCC   CUGAUGAG   X   CGAA   ACGCAGG   CCUGCGU   C   GGCACGU       331   HCV-6106   6106   CGAAGCG   CUGAUGAG   X   CGAA   ACGCUAU   AUAGCGU   U   CGCUUCG       332   HCV-6107   6107   GCGAAGC   CUGAUGAG   X   CGAA   AACGCUA   UAGCGUU   C   GCUUCGC       333   HCV-6111   6111   CCCCGCG   CUGAUGAG   X   CGAA   AGCGAAC   GUUCGCU   U   CGCGGGG       334   HCV-6413   6413   UUUGCAU   CUGAUGAG   X   CGAA   AUGCCGU   ACGGCAU   C   AUGCAAA       335   HCV-6574   6574   CCUGGAA   CUGAUGAG   X   CGAA   AGUUCGG   CCGAACU   A   UUCCAGG       336   HCV-6576   6576   GCCCUGG   CUGAUGAG   X   CGAA   AUAGUUC   GAACUAU   U   CCAGGGC       337   HCV-6577   6577   CGCCCUG   CUGAUGAG   X   CGAA   AAUAGUU   AACUAUU   C   CAGGGCG       338   HCV-6637   6637   GUAGUGG   CUGAUGAG   X   CGAA   AGUCCCC   GGGGACU   U   CCACUAC       339   HCV-6638   6638   CGUAGUG   CUGAUGAG   X   CGAA   AAGUCCC   GGGACUU   C   CACUACG       340   HCV-6643   6643   CGUCACG   CUGAUGAG   X   CGAA   AGUGGAA   UUCCACU   A   CGUGACG       341   HCV-6671   6671   GGCAUUU   CUGAUGAG   X   CGAA   ACGUUGU   ACAACGU   A   AAAUGCC       342   HCV-6703   6703   GGUGAAG   CUGAUGAG   X   CGAA   AUUCGGG   CCCGAAU   U   CUUCACC       343   HCV-6704   6704   CGGUGAA   CUGAUGAG   X   CGAA   AAUUCGG   CCGAAUU   C   UUCACCG       344   HCV-6706   6706   UUCGGUG   CUGAUGAG   X   CGAA   AGAAUUC   GAAUUCU   U   CACCGAA       345   HCV-6707   6707   AUUCGGU   CUGAUGAG   X   CGAA   AAGAAUU   AAUUCUU   C   ACCGAAU       346   HCV-6715   6715   CCCGUCC   CUGAUGAG   X   CGAA   AUUCGGU   ACCGAAU   U   GGACGGG       347   HCV-6730   6730   CCUGUGC   CUGAUGAG   X   CGAA   ACCGCAC   GUGCGGU   U   GCACAGG       348   HCV-6739   6739   CGGAGCG   CUGAUGAG   X   CGAA   ACCUGUG   CACAGGU   A   CGCUCCG       349   HCV-6744   6744   CACGCCG   CUGAUGAG   X   CGAA   AGCGUAC   GUACGCU   C   CGGCGUG       350   HCV-6759   6759   CGUAGGA   CUGAUGAG   X   CGAA   AGGUCUG   CAGACCU   C   UCCUACG       351   HCV-6761   6761   CCCGUAG   CUGAUGAG   X   CGAA   AGAGGUC   GACCUCU   C   CUACGGG       352   HCV-6764   6764   CCUCCCG   CUGAUGAG   X   CGAA   AGGAGAG   CUCUCCU   A   CGGGAGG       353   HCV-6776   6776   GGAAUGU   CUGAUGAG   X   CGAA   ACAUCCU   AGGAUGU   C   ACAUUCC       354   HCV-6782   6782   CGACCUG   CUGAUGAG   X   CGAA   AAUGUGA   UCACAUU   C   CAGGUCG       355   HCV-6788   6788   UGAGCCC   CUGAUGAG   X   CGAA   ACCUGGA   UCCAGGU   C   GGGCUCA       356   HCV-6794   6794   AUUGGUU   CUGAUGAG   X   CGAA   AGCCCGA   UCGGGCU   C   AACCAAU       357   HCV-6802   6802   AACCAGG   CUGAUGAG   X   CGAA   AUUGGUU   AACCAAU   A   CCUGGUU       358   HCV-6809   6809   GUGACCC   CUGAUGAG   X   CGAA   ACCAGGU   ACCUGGU   U   GGGUCAC       359   HCV-6814   6814   GAGCUGU   CUGAUGAG   X   CGAA   ACCCAAC   GUUGGGU   C   ACAGCUC       360   HCV-6821   6821   CGCAUGG   CUGAUGAG   X   CGAA   AGCUGUG   CACAGCU   C   CCAUGCG       361   HCV-6906   6906   GCCAGCC   CUGAUGAG   X   CGAA   ACGUUUA   UAAACGU   A   GGCUGGC       362   HCV-6922   6922   GGGGGGA   CUGAUGAG   X   CGAA   ACCCCCU   AGGGGGU   C   UCCCCCC       363   HCV-6924   6924   GAGGGGG   CUGAUGAG   X   CGAA   AGACCCC   GGGGUCU   C   CCCCCUC       364   HCV-6931   6931   GGCCAAG   CUGAUGAG   X   CGAA   AGGGGGG   CCCCCCU   C   CUUGGCC       365   HCV-6934   6934   GCUGGCC   CUGAUGAG   X   CGAA   AGGAGGG   CCCUCCU   U   GGCCAGC       366   HCV-6943   6943   AGCUGAA   CUGAUGAG   X   CGAA   AGCUGGC   GCCAGCU   C   UUCAGCU       367   HCV-6958   6958   CGCAGAC   CUGAUGAG   X   CGAA   AUUGGCU   AGCCAAU   U   GUCUGCG       368   HCV-6961   6961   AGGCGCA   CUGAUGAG   X   CGAA   ACAAUUG   CAAUUGU   C   UGCGCCU       369   HCV-7034   7034   GCCACAG   CUGAUGAG   X   CGAA   AGGUUGG   CCAACCU   C   CUGUGGC       370   HCV-7118   7118   CCGCUCG   CUGAUGAG   X   CGAA   AGCGGGU   ACCCGCU   U   CGAGCGG       371   HCV-7119   7119   UCCGCUC   CUGAUGAG   X   CGAA   AAGCGGG   CCCGCUU   C   GAGCGGA       372   HCV-7145   7145   CAACGGA   CUGAUGAG   X   CGAA   ACUUCCC   GGGAAGU   A   UCCGUUG       373   HCV-7195   7195   UAUGGGC   CUGAUGAG   X   CGAA   ACGCGGG   CCCGCGU   U   GCCCAUA       374   HCV-7202   7202   GUGCCCA   CUGAUGAG   X   CGAA   AUGGGCA   UGCCCAU   A   UGGGCAC       375   HCV-7218   7213   GGGUUGU   CUGAUGAG   X   CGAA   AUCCGGG   CCCGGAU   U   ACAACCC       376   HCV-7219   7219   AGGGUUG   CUGAUGAG   X   CGAA   AAUCCGG   CCGGAUU   A   CAACCCU       377   HCV-7234   7234   GGACUCU   CUGAUGAG   X   CGAA   ACAGUGG   CCACUGU   U   AGAGUCC       378   HCV-7235   7235   AGGACUC   CUGAUGAG   X   CGAA   AACAGUG   CACUGUU   A   GAGUCCU       379   HCV-7251   7251   UAGUCCG   CUGAUGAG   X   CGAA   ACUUUUC   GAAAAGU   C   CGGACUA       380   HCV-7258   7258   AGGGACG   CUGAUGAG   X   CGAA   AGUCCGG   CCGGACU   A   CGUCCCU       381   HCV-7262   7262   CCGGAGG   CUGAUGAG   X   CGAA   ACGUAGU   ACUACGU   C   CCUCCGG       382   HCV-7266   7266   ACCGCCG   CUGAUGAG   X   CGAA   AGGGACG   CGUCCCU   C   CGGCGGU       383   HCV-7288   7288   AGGCGGC   CUGAUGAG   X   CGAA   AUGGGCA   UGCCCAU   U   GCCGCCU       384   HCV-7296   7296   CCCGUGG   CUGAUGAG   X   CGAA   AGGCGGC   GCCGCCU   A   CCACGGG       385   HCV-7354   7354   CACGGUG   CUGAUGAG   X   CGAA   ACUCUGU   ACAGAGU   C   CACCGUG       386   HCV-7386   7386   GUCUUAG   CUGAUGAG   X   CGAA   AGCCAGC   GCUGGCU   A   CUAAGAC       387   HCV-7389   7389   AAAGUCU   CUGAUGAG   X   CGAA   AGUAGCC   G3CUACU   A   AGACUUU       388   HCV-7395   7395   CUGCCGA   CUGAUGAG   X   CGAA   AGUCUUA   UAAGACU   U   UCGGCAG       389   HCV-7396   7396   GCUGCCG   CUGAUGAG   X   CGAA   AAGUCUU   AAGACUU   U   CGGCAGC       390   HCV-7397   7397   AGCUGCC   CUGAUGAG   X   CGAA   AAAGUCU   AGACUUU   C   GGCAGCU       391   HCV-7411   7411   GCCCGAC   CUGAUGAG   X   CGAA   AUCCGGA   UCCGGAU   C   GUCGGCC       392   HCV-7414   7414   AACGGCC   CUGAUGAG   X   CGAA   ACGAUCC   GGAUCGU   C   GGCCGUU       393   HCV-7421   7421   CGCUGUC   CUGAUGAG   X   CGAA   ACGGCCG   CGGCCGU   U   GACAGCG       394   HCV-7498   7498   CAUGGAG   CUGAUGAG   X   CGAA   AGUACGA   UCGUACU   C   CUCCAUG       395   HCV-7501   7501   GGGCAUG   CUGAUGAG   X   CGAA   AGGAGUA   UACUCCU   C   CAUGCCC       396   HCV-7514   7514   CCCCCUC   CUGAUGAG   X   CGAA   AGGGGGG   CCCCCCU   U   GAGGGGG       397   HCV-7539   7539   UCGCUGA   CUGAUGAG   X   CGAA   AUCAGGG   CCCUGAU   C   UCAGCGA       398   HCV-7541   7541   CGUCGCU   CUGAUGAG   X   CGAA   AGAUCAG   CUGAUCU   C   AGCGACG       399   HCV-7552   7552   AGACCAA   CUGAUGAG   X   CGAA   ACCCGUC   GACGGGU   C   UUGGUCU       400   HCV-7554   7554   GUAGACC   CUGAUGAG   X   CGAA   AGACCCG   CGGGUCU   U   GGUCUAC       401   HCV-7558   7558   CACGGUA   CUGAUGAG   X   CGAA   ACCAAGA   UCUUGGU   C   UACCGUG       402   HCV-7560   7560   CUCACGG   CUGAUGAG   X   CGAA   AGACCAA   UUGGUCU   A   CCGUGAG       403   HCV-7589   7589   AGCAGAC   CUGAUGAG   X   CGAA   AUGUCGU   ACGACAU   C   GUCUGCU       404   HCV-7592   7592   AGCAGCA   CUGAUGAG   X   CGAA   ACGAUGU   ACAUCGU   C   UGCUGCU       405   HCV-7600   7600   GGACAUU   CUGAUGAG   X   CGAA   AGCAGCA   UGCUGCU   C   AAUGUCC       406   HCV-7606   7606   UGUGUAG   CUGAUGAG   X   CGAA   ACAUUGA   UCAAUGU   C   CUACACA       407   HCV-7667   7667   ACGCGUU   CUGAUGAG   X   CGAA   AUGGGCA   UGCCCAU   C   AACGCGU       408   HCV-7723   7723   ACUGCGG   CUGAUGAG   X   CGAA   AUGUUGU   ACAACAU   C   CCGCAGU       409   HCV-7775   7775   CGUCCAG   CUGAUGAG   X   CGAA   ACUUGCA   UGCAAGU   C   CUGGACG       410   HCV-7789   7789   GUCCCGG   CUGAUGAG   X   CGAA   AGUGGUC   GACCACU   A   CCGGGAC       411   HCV-7839   7839   AGAAGUU   CUGAUGAG   X   CGAA   AGCCUUA   UAAGGCU   A   AACUUCU       412   HCV-7847   7847   CUACGGA   CUGAUGAG   X   CGAA   AGAAGUU   AACUUCU   A   UCCGUAG       413   HCV-7849   7849   UUCUACG   CUGAUGAG   X   CGAA   AUAGAAG   CUUCUAU   C   CGUAGAA       414   HCV-7853   7853   CUUCUUC   CUGAUGAG   X   CGAA   ACGGAUA   UAUCCGU   A   GAAGAAG       415   HCV-7894   7894   AAAUUUA   CUGAUGAG   X   CGAA   AUUUGGC   GCCAAAU   C   UAAAUUU       416   HCV-7896   7896   CCAAAUU   CUGAUGAG   X   CGAA   AGAUUUG   CAAAUCU   A   AAUUUGG       417   HCV-7900   7900   AUAGCCA   CUGAUGAG   X   CGAA   AUUUAGA   UCUAAAU   U   UGGCUAU       418   HCV-7901   7901   CAUAGCC   CUGAUGAG   X   CGAA   AAUUUAG   CUAAAUU   U   GGCUAUG       419   HCV-7906   7906   UGCCCCA   CUGAUGAG   X   CGAA   AGCCAAA   UUUGGCU   A   UGGGGCA       420   HCV-7955   7955   CGGAGCG   CUGAUGAG   X   CGAA   AUGUGGU   ACCACAU   C   CGCUCCG       421   HCV-7960   7960   CCACACG   CUGAUGAG   X   CGAA   AGCGGAU   AUCCGCU   C   CGUGUGG       422   HCV-8075   8075   AUACGAU   CUGAUGAG   X   CGAA   AGGCGAG   CUCGCCU   U   AUCGUAU       423   HCV-8076   8076   AAUACGA   CUGAUGAG   X   CGAA   AAGGCGA   UCGCCUU   A   UCGUAUU       424   HCV-8078   8078   GGAAUAC   CUGAUGAG   X   CGAA   AUAAGGC   GCCUUAU   C   GUAUUCC       425   HCV-8170   8170   GAAUCCG   CUGAUGAG   X   CGAA   ACGAGGA   UCCUCGU   A   CGGAUUC       426   HCV-8176   8176   GUACUGG   CUGAUGAG   X   CGAA   AUCCGUA   UACGGAU   U   CCAGUAC       427   HCV-8182   8182   AGGAGAG   CUGAUGAG   X   CGAA   ACUGGAA   UUCCAGU   A   CUCUCCU       428   HCV-8187   8187   UGCCCAG   CUGAUGAG   X   CGAA   AGAGUAC   GUACUCU   C   CUGGGCA       429   HCV-8201   8201   GGAACUC   CUGAUGAG   X   CGAA   ACCCGCU   AGCGGGU   U   GAGUUCC       430   HCV-8206   8206   CACCAGG   CUGAUGAG   X   CGAA   ACUCAAC   GUUGAGU   U   CCUGGUG       431   HCV-8207   8207   UCACCAG   CUGAUGAG   X   CGAA   AACUCAA   UUGAGUU   C   CUGGUGA       432   HCV-8227   8227   UUUCUUU   CUGAUGAG   X   CGAA   AUUUCCA   UGGAAAU   C   AAAGAAA       433   HCV-8357   8357   GCGACUU   CUGAUGAG   X   CGAA   AUGGCCU   AGGCCAU   A   AAGUCGC       434   HCV-8362   8362   CGUGAGC   CUGAUGAG   X   CGAA   ACUUUAU   AUAAAGU   C   GCUCACG       435   HCV-8366   8366   GCUCCGU   CUGAUGAG   X   CGAA   AGCGACU   AGUCGCU   C   ACGGAGC       436   HCV-8378   8378   CGAUGUA   CUGAUGAG   X   CGAA   AGCCGCU   AGCGGCU   C   UACAUCG       437   HCV-8380   8380   CCCGAUG   CUGAUGAG   X   CGAA   AGAGCCG   CGGCUCU   A   CAUCGGG       438   HCV-8384   8384   GGCCCCC   CUGAUGAG   X   CGAA   AUGUAGA   UCUACAU   C   GGGGGCC       439   HCV-8424   8424   CGGCGAU   CUGAUGAG   X   CGAA   ACCGCAG   CUGCGGU   U   AUCGCCG       440   HCV-8425   8425   CCGGCGA   CUGAUGAG   X   CGAA   AACCGCA   UGCGGUU   A   UCGCCGG       441   HCV-8427   8427   CACCGGC   CUGAUGAG   X   CGAA   AUAACCG   CGGUUAU   C   GCCGGUG       442   HCV-8460   8460   CCGCAGC   CUGAUGAG   X   CGAA   AGUCGUC   GACGACU   A   GCUGCGG       443   HCV-8508   8508   GCAGCUC   CUGAUGAG   X   CGAA   ACAGGCC   GGCCUGU   C   GAGCUGC       444   HCV-8522   8522   AGUCCUG   CUGAUGAG   X   CGAA   AGCUUUG   CAAAGCU   C   CAGGACU       445   HCV-8540   8540   CGUUCAC   CUGAUGAG   X   CGAA   AGCAUCG   CGAUGCU   C   GUGAACG       446   HCV-8558   8558   UAACGAC   CUGAUGAG   X   CGAA   AGGUCGU   ACGACCU   U   GUCGUUA       447   HCV-8561   8561   AGAUAAC   CUGAUGAG   X   CGAA   ACAAGGU   ACCUUGU   C   GUUAUCU       448   HCV-8564   8564   CACAGAU   CUGAUGAG   X   CGAA   ACGACAA   UUGUCGU   U   AUCUGUG       449   HCV-8638   8638   GGGGGCA   CUGAUGAG   X   CGAA   AGUACCU   AGGUACU   C   UGCCCCC       450   HCV-8671   8671   CAAGUCG   CUGAUGAG   X   CGAA   AUUCUGG   CCAGAAU   A   CGACUUG       451   HCV-8698   8698   GUUGGAG   CUGAUGAG   X   CGAA   AGCAUGA   UCAUGCU   C   CUCCAAC       452   HCV-8701   8701   CACGUUG   CUGAUGAG   X   CGAA   AGGAGCA   UGCUCCU   C   CAACGUG       453   HCV-8728   8728   UUUGCCG   CUGAUGAG   X   CGAA   AUGCGUC   GACGCAU   C   CGGCAAA       454   HCV-8774   8774   CCCGUGC   CUGAUGAG   X   CGAA   AGGGGGG   CCCCCCU   U   GCACGGG       455   HCV-8842   8842   GGGCGCA   CUGAUGAG   X   CGAA   ACAUGAU   AUCAUGU   A   UGCGCCC       456   HCV-8854   8854   UGCCCAU   CUGAUGAG   X   CGAA   AGGUGGG   CCCACCU   U   AUGGGCA       457   HCV-8855   8855   UUGCCCA   CUGAUGAG   X   CGAA   AAGGUGG   CCACCUU   A   UGGGCAA       458   HCV-8871   8871   GUCAUCA   CUGAUGAG   X   CGAA   AAUCAUC   GAUGAUU   U   UGAUGAC       459   HCV-8880   8880   AAGAAGU   CUGAUGAG   X   CGAA   AGUCAUC   GAUGACU   C   ACUUCUU       460   HCV-8931   8931   AUCUGAC   CUGAUGAG   X   CGAA   AUCCAGG   CCUGGAU   U   GUCAGAU       461   HCV-8934   8934   UAGAUCU   CUGAUGAG   X   CGAA   ACAAUCC   GGAUUGU   C   AGAUCUA       462   HCV-8939   8939   CCCCGUA   CUGAUGAG   X   CGAA   AUCUGAC   GUCAGAU   C   UACGGGG       463   HCV-8941   8941   GGCCCCG   CUGAUGAG   X   CGAA   AGAUCUG   CAGAUCU   A   CGGGGCC       464   HCV-9065   9065   GUUUCCU   CUGAUGAG   X   CGAA   AGGCAUG   CAUGCCU   C   AGGAAAC       465   HCV-9074   9074   GUACCCC   CUGAUGAG   X   CGAA   AGUUUCC   GGAAACU   U   GGGGUAC       466   HCV-9080   9080   AGGGCGG   CUGAUGAG   X   CGAA   ACCCCAA   UUGGGGU   A   CCGCCCU       467   HCV-9088   9088   GACUCGC   CUGAUGAG   X   CGAA   AGGGCGG   CCGCCCU   U   GCGAGUC       468   HCV-9095   9095   GUCUCCA   CUGAUGAG   X   CGAA   ACUCGCA   UGCGAGU   C   UGGAGAC       469   HCV-9119   9119   UAGCGCG   CUGAUGAG   X   CGAA   ACACUUC   GAAGUGU   C   CGCGCUA       470   HCV-9126   9126   AGUAGCC   CUGAUGAG   X   CGAA   AGCGCGG   CCGCGCU   A   GGCUACU       471   HCV-9131   9131   GGGACAG   CUGAUGAG   X   CGAA   AGCCUAG   CUAGGCU   A   CUGUCCC       472   HCV-9136   9136   CCCUUGG   CUGAUGAG   X   CGAA   ACAGUAG   CUACUGU   C   CCAAGGG       473   HCV-9226   9226   CAGCUGG   CUGAUGAG   X   CGAA   ACGCGGC   GCCGCGU   C   CCAGCUG       474   HCV-9238   9238   GCUGGAC   CUGAUGAG   X   CGAA   AGUCCAG   CUGGACU   U   GUCCAGC       475   HCV-9241   9241   CCAGCUG   CUGAUGAG   X   CGAA   ACAAGUC   GACUUGU   C   CAGCUGG       476   HCV-9250   9250   AGCAACG   CUGAUGAG   X   CGAA   ACCAGCU   AGCUGGU   U   CGUUGCU       477   HCV-9251   9251   CAGCAAC   CUGAUGAG   X   CGAA   AACCAGC   GCUGGUU   C   GUUGCUG       478   HCV-9254   9254   AACCAGC   CUGAUGAG   X   CGAA   ACGAACC   GGUUCGU   U   GCUGGUU       479   HCV-9278   9278   UGUGAUA   CUGAUGAG   X   CGAA   AUGUCUC   GAGACAU   A   UAUCACA       480   HCV-9280   9280   GCUGUGA   CUGAUGAG   X   CGAA   AUAUGUC   GACAUAU   A   UCACAGC       481   HCV-9282   9282   AGGCUGU   CUGAUGAG   X   CGAA   AUAUAUG   CAUAUAU   C   ACAGCCU       482   HCV-9292   9292   GGCACGA   CUGAUGAG   X   CGAA   ACAGGCU   AGCCUGU   C   UCGUGCC       483   HCV-9326   9326   GUAGGAG   CUGAUGAG   X   CGAA   AGGCACC   GGUGCCU   A   CUCCUAC       484   HCV-9329   9329   AAAGUAG   CUGAUGAG   X   CGAA   AGUAGGC   GCCUACU   C   CUACUUU       485   HCV-9332   9332   CGGAAAG   CUGAUGAG   X   CGAA   AGGAGUA   UACUCCU   A   CUUUCCG       486   HCV-9335   9335   CUACGGA   CUGAUGAG   X   CGAA   AGUAGGA   UCCUACU   U   UCCGUAG       487   HCV-9336   9336   CCUACGG   CUGAUGAG   X   CGAA   AAGUAGG   CCUACUU   U   CCGUAGG       488   HCV-9337   9337   CCCUACG   CUGAUGAG   X   CGAA   AAAGUAG   CUACUUU   C   CGUAGGG       489   HCV-9341   9341   CUACCCC   CUGAUGAG   X   CGAA   ACGGAAA   UUUCCGU   A   GGGGUAG       490   HCV-9347   9347   AGAUGCC   CUGAUGAG   X   CGAA   ACCCCUA   UAGGGGU   A   GGCAUCU       491   HCV-9353   9353   GCAGGUA   CUGAUGAG   X   CGAA   AUGCCUA   UAGGCAU   C   UACCUGC       492   HCV-9355   9355   GAGCAGG   CUGAUGAG   X   CGAA   AGAUGCC   GGCAUCU   A   CCUGCUC       493   HCV-9362   9362   GGUUGGG   CUGAUGAG   X   CGAA   AGCAGGU   ACCUGCU   C   CCCAACC       494   HCV-9385   9385   GAGUGAU   CUGAUGAG   X   CGAA   AGCUCCC   GGGAGCU   A   AUCACUC       495   HCV-9388   9388   CUGGAGU   CUGAUGAG   X   CGAA   AUUAGCU   AGCUAAU   C   ACUCCAG       496   HCV-9392   9392   UGGCCUG   CUGAUGAG   X   CGAA   AGUGAUU   AAUCACU   C   CAGGCCA       497   HCV-9402   9402   GAUGGCC   CUGAUGAG   X   CGAA   AUUGGCC   GGCCAAU   A   GGCCAUC                    
     [0298] Where “X” represents stem II region of a HH ribozyme (Hertel et al., 1992  Nucleic Acids Res.  20:3252). The length of stem II may be 2 base-pairs.  
               TABLE VI                          Additional HCV Hammerhead (HH) Ribozyme and Target Sequence                         Pos.   Ribozyme   Substrate                                                         14   CGCCCCC   CUGAUGAG   X   CGAA   AUCGGGG   CCCCGAU   U   GGGGGCG       34   AGUGAUC   CUGAUGAG   X   CGAA   AUGGUGG   CCACCAU   A   GAUCACU       38   GGGGAGU   CUGAUGAG   X   CGAA   AUCUAUG   CAUAGAU   C   ACUCCCC       42   CACAGGG   CUGAUGAG   X   CGAA   AGUGAUC   GAUCACU   C   CCCUGUG       57   AAGACAG   CUGAUGAG   X   CGAA   AGUUCCU   AGGAACU   A   CUGUCUU       62   GCGUGAA   CUGAUGAG   X   CGAA   ACAGUAG   CUACUGU   C   UUCACGC       64   CUGCGUG   CUGAUGAG   X   CGAA   AGACAGU   ACUGUCU   U   CACGCAG       65   UCUGCGU   CUGAUGAG   X   CGAA   AAGACAG   CUGUCUU   C   ACGCAGA       79   AUGGCUA   CUGAUGAG   X   CGAA   ACGCUUU   AAAGCGU   C   UAGCCAU       81   CCAUGGC   CUGAUGAG   X   CGAA   AGACGCU   AGCGUCU   A   GCCAUGG       92   UCAUACU   CUGAUGAG   X   CGAA   ACGCCAU   AUGGCGU   U   AGUAUGA       93   CUCAUAC   CUGAUGAG   X   CGAA   AACGCCA   UGGCGUU   A   GUAUGAG       96   ACACUCA   CUGAUGAG   X   CGAA   ACUAACG   CGUUAGU   A   UGAGUGU       104   GCUGCAC   CUGAUGAG   X   CGAA   ACACUCA   UGAGUGU   C   GUGCAGC       142   AGACCAC   CUGAUGAG   X   CGAA   AUGGCUC   GAGCCAU   A   GUGGUCU       192   AAGAAAG   CUGAUGAG   X   CGAA   ACCCGGU   ACCGGGU   C   CUUUCUU       195   UCCAAGA   CUGAUGAG   X   CGAA   AGGACCC   GGGUCCU   U   UCUUGGA       196   AUCCAAG   CUGAUGAG   X   CGAA   AAGGACC   GGUCCUU   U   CUUGGAU       197   GAUCCAA   CUGAUGAG   X   CGAA   AAAGGAC   GUCCUUU   C   UUGGAUC       204   GCGGGUU   CUGAUGAG   X   CGAA   AUCCAAG   CUUGGAU   C   AACCCGC       227   ACGCCCA   CUGAUGAG   X   CGAA   AUCUCCA   UGGAGAU   U   UGGGCGU       228   CACGCCC   CUGAUGAG   X   CGAA   AAUCUCC   GGAGAUU   U   GGGCGUG       282   GUACCAC   CUGAUGAG   X   CGAA   AGGCCUU   AAGGCCU   U   GUGGUAC       354   GGUUUAG   CUGAUGAG   X   CGAA   AUUCGUG   CACGAAU   C   CUAAACC       357   UGAGGUU   CUGAUGAG   X   CGAA   AGGAUUC   GAAUCCU   A   AACCUCA       363   UUUCUUU   CUGAUGAG   X   CGAA   AGGUUUA   UAAACCU   C   AAAGAAA       381   UAGGUGU   CUGAUGAG   X   CGAA   ACGUUUG   CAAACGU   A   ACACCUA       388   GCGGCGG   CUGAUGAG   X   CGAA   AGGUGUU   AACACCU   A   CCGCCGC       431   CACCAAC   CUGAUGAG   X   CGAA   AUCUGAC   GUCAGAU   C   GUUGGUG       434   CUCCACC   CUGAUGAG   X   CGAA   ACGAUCU   AGAUCGU   U   GGUGGAG       443   ACACGUA   CUGAUGAG   X   CGAA   ACUCCAC   GUGGAGU   U   UACGUGU       444   AACACGU   CUGAUGAG   X   CGAA   AACUCCA   UGGAGUU   U   ACGUGUU       445   CAACACG   CUGAUGAG   X   CGAA   AAACUCC   GGAGUUU   A   CGUGUUG       451   GCGCGGC   CUGAUGAG   X   CGAA   ACACGUA   UACGUGU   U   GCCGCGC       516   CUUCCAC   CUGAUGAG   X   CGAA   AGGUUGC   GCAACCU   C   GUGGAAG       688   AUUGCGC   CUGAUGAG   X   CGAA   ACCUCCG   CGGAGGU   C   GCGCAAU       702   AUGACCU   CUGAUGAG   X   CGAA   ACCCAGA   UCUGGGU   A   AGGUCAU       719   CGCACGU   CUGAUGAG   X   CGAA   AGGGUAU   AUACCCU   C   ACGUGCG       740   ACCCCAU   CUGAUGAG   X   CGAA   AGGUCGG   CCGACCU   C   AUGGGGU       861   AUAGAGA   CUGAUGAG   X   CGAA   AGAGCAA   UUGCUCU   U   UCUCUAU       862   GAUAGAG   CUGAUGAG   X   CGAA   AAGAGCA   UGCUCUU   U   CUCUAUC       863   AGAUAGA   CUGAUGAG   X   CGAA   AAAGAGC   GCUCUUU   C   UCUAUCU       865   GAAGAUA   CUGAUGAG   X   CGAA   AGAAAGA   UCUUUCU   C   UAUCUUC       867   AGGAAGA   CUGAUGAG   X   CGAA   AGAGAAA   UUUCUCU   A   UCUUCCU       869   AGAGGAA   CUGAUGAG   X   CGAA   AUAGAGA   UCUCUAU   C   UUCCUCU       871   CAAGAGG   CUGAUGAG   X   CGAA   AGAUAGA   UCUAUCU   U   CCUCUUG       872   CCAAGAG   CUGAUGAG   X   CGAA   AAGAUAG   CUAUCUU   C   CUCUUGG       875   GGGCCAA   CUGAUGAG   X   CGAA   AGGAAGA   UCUUCCU   C   UUGGCCC       877   CAGGGCC   CUGAUGAG   X   CGAA   AGAGGAA   UUCCUCU   U   GGCCCUG       889   CAAACAG   CUGAUGAG   X   CGAA   ACAGCAG   CUGCUGU   C   CUGUUUG       894   AUGGUCA   CUGAUGAG   X   CGAA   ACAGGAC   GUCCUGU   U   UGACCAU       895   GAUGGUC   CUGAUGAG   X   CGAA   AACAGGA   UCCUGUU   U   GACCAUC       902   AAGCUGG   CUGAUGAG   X   CGAA   AUGGUCA   UGACCAU   C   CCAGCUU       909   UAAGCGG   CUGAUGAG   X   CGAA   AGCUGGG   CCCAGCU   U   CCGCUUA       910   AUAAGCG   CUGAUGAG   X   CGAA   AAGCUGG   CCAGCUU   C   CGCUUAU       915   ACCUGAU   CUGAUGAG   X   CGAA   AGCGGAA   UUCCGCU   U   AUCAGGU       916   CACCUGA   CUGAUGAG   X   CGAA   AAGCGGA   UCCGCUU   A   UCAGGUG       918   CGCACCU   CUGAUGAG   X   CGAA   AUAAGCG   CGCUUAU   C   AGGUGCG       934   CAGCCCG   CUGAUGAG   X   CGAA   AUGCGUU   AACGCAU   C   CGGGCUG       943   GACAUGG   CUGAUGAG   X   CGAA   ACAGCCC   GGGCUGU   A   CCAUGUC       950   CAUUCGU   CUGAUGAG   X   CGAA   ACAUGGU   ACCAUGU   C   ACGAAUG       964   UGAGUUG   CUGAUGAG   X   CGAA   AGCAGUC   GACUGCU   C   CAACUCA       970   AAUGCUU   CUGAUGAG   X   CGAA   AGUUGGA   UCCAACU   C   AAGCAUU       977   CAUACAC   CUGAUGAG   X   CGAA   AUGCUUG   CAAGCAU   U   GUGUAUG       1008   CCGGGGG   CUGAUGAG   X   CGAA   AUGCAUG   CAUGCAU   A   CCCCCGG       1067   UGGGAGU   CUGAUGAG   X   CGAA   AGCGCUA   UAGCGCU   C   ACUCCCA       1071   AGCGUGG   CUGAUGAG   X   CGAA   AGUGAGC   GCUCACU   C   CCACGCU       1079   UGGCCGC   CUGAUGAG   X   CGAA   AGCGUGG   CCACGCU   C   GCGGCCA       1100   UAGUGGG   CUGAUGAG   X   CGAA   AUGCUGG   CCAGCAU   C   CCCACUA       1107   AUUGUCG   CUGAUGAG   X   CGAA   AGUGGGG   CCCCACU   A   CGACAAU       1115   GGCGUCG   CUGAUGAG   X   CGAA   AUUGUCG   CGACAAU   A   CGACGCC       1152   GAACAGA   CUGAUGAG   X   CGAA   AGCGGCC   GGCCGCU   U   UCUGUUC       1181   AUCCGCA   CUGAUGAG   X   CGAA   AGGUCCC   GGGACCU   C   UGCGGAU       1199   GGGAGAC   CUGAUGAG   X   CGAA   AGGAAAA   UUUUCCU   C   GUCUCCC       1202   ACUGGGA   CUGAUGAG   X   CGAA   ACGAGGA   UCCUCGU   C   UCCCAGU       1204   CAACUGG   CUGAUGAG   X   CGAA   AGACGAG   CUCGUCU   C   CCAGUUG       1210   GGUGAAC   CUGAUGAG   X   CGAA   ACUGGGA   UCCCAGU   U   GUUCACC       1213   GAAGGUG   CUGAUGAG   X   CGAA   ACAACUG   CAGUUGU   U   CACCUUC       1214   AGAAGGU   CUGAUGAG   X   CGAA   AACAACU   AGUUGUU   C   ACCUUCU       1219   AGGCGAG   CUGAUGAG   X   CGAA   AGGUGAA   UUCACCU   U   CUCGCCU       1220   GAGGCGA   CUGAUGAG   X   CGAA   AAGGUGA   UCACCUU   C   UCGCCUC       1222   GCGAGGC   CUGAUGAG   X   CGAA   AGAAGGU   ACCUUCU   C   GCCUCGC       1227   UACCGGC   CUGAUGAG   X   CGAA   AGGCGAG   CUCGCCU   C   GCCGGUA       1234   UGUCUCA   CUGAUGAG   X   CGAA   ACCGGCG   CGCCGGU   A   UGAGACA       1244   AGUCCUG   CUGAUGAG   X   CGAA   ACUGUCU   AGACAGU   A   CAGGACU       1257   AUUGAGC   CUGAUGAG   X   CGAA   AUUGCAG   CUGCAAU   U   GCUCAAU       1261   AUAGAUU   CUGAUGAG   X   CGAA   AGCAAUU   AAUUGCU   C   AAUCUAU       1265   CGGGAUA   CUGAUGAG   X   CGAA   AUUGAGC   GCUCAAU   C   UAUCCCG       1267   GCCGGGA   CUGAUGAG   X   CGAA   AGAUGGA   UCAAUCU   A   UCCCGGC       1269   UGGCCGG   CUGAUGAG   X   CGAA   AUAGAUU   AAUCUAU   C   CCGGCCA       1299   AUAUCCC   CUGAUGAG   X   CGAA   AGCCAUG   CAUGGCU   U   GGGAUAU       1305   AUCAUCA   CUGAUGAG   X   CGAA   AUCCCAA   UUGGGAU   A   UGAUGAU       1321   UGUAGGC   CUGAUGAG   X   CGAA   ACCAGUU   AACUGGU   C   GCCUACA       1326   GCUGUUG   CUGAUGAG   X   CGAA   AGGCGAC   GUCGCCU   A   CAACAGC       1337   ACACCAC   CUGAUGAG   X   CGAA   AGGGCUG   CAGCCCU   A   GUGGUGU       1345   UAACUGC   CUGAUGAG   X   CGAA   ACACCAC   GUGGUGU   C   GCAGUUA       1351   CCGGAGU   CUGAUGAG   X   CGAA   ACUGCGA   UCGCAGU   U   ACUCCGG       1352   UCCGGAG   CUGAUGAG   X   CGAA   AACUGCG   CGCAGUU   A   CUCCGGA       1355   GGAUCCG   CUGAUGAG   X   CGAA   AGUAACU   AGUUACU   C   CGGAUCC       1361   CUGGUGG   CUGAUGAG   X   CGAA   AUCCGGA   UCCGGAU   C   CCACAAG       1449   AAGACCU   CUGAUGAG   X   CGAA   AGCCCAG   CUGGGCU   A   AGGUCUU       1454   CAAUCAA   CUGAUGAG   X   CGAA   ACCUUAG   CUAAGGU   C   UUGAUUG       1456   CACAAUC   CUGAUGAG   X   CGAA   AGACCUC   AAGGUCU   U   GAUUGUG       1460   ACAUCAC   CUGAUGAG   X   CGAA   AUCAAGA   UCUUGAU   U   GUGAUGU       1468   AAAGAGU   CUGAUGAG   X   CGAA   ACAUCAC   GUGAUGU   U   ACUCUUU       1469   CAAAGAG   CUGAUGAG   X   CGAA   AACAUCA   UGAUGUU   A   CUCUUUG       1472   CGGCAAA   CUGAUGAG   X   CGAA   AGUAACA   UGUUACU   C   UUUGCCG       1474   GCCGGCA   CUGAUGAG   X   CGAA   AGAGUAA   UUACUCU   U   UGCCGGC       1475   CGCCGGC   CUGAUGAG   X   CGAA   AAGAGUA   UACUCUU   U   GCCGGCG       1484   CCCCGUC   CUGAUGAG   X   CGAA   ACGCCGG   CCGGCGU   U   GACGGGG       1493   UGUAAGU   CUGAUGAG   X   CGAA   ACCCCGU   ACGGGGU   C   ACUUACA       1497   GUCGUGU   CUGAUGAG   X   CGAA   AGUGACC   GGUCACU   U   ACACGAC       1498   UGUCGUG   CUGAUGAG   X   CGAA   AAGUGAC   GUCACUG   A   CACGACA       1513   AGCUUGC   CUGAUGAG   X   CGAA   ACCCCCC   GGGGGGU   C   GCAAGCU       1521   GUGUGGC   CUGAUGAG   X   CGAA   AGCUUGC   GCAAGCU   C   GCCACAC       1538   AGGACGU   CUGAUGAG   X   CGAA   ACGCUCU   AGAGCGU   C   ACGUCCU       1543   GAAGAAG   CUGAUGAG   X   CGAA   ACGUGAC   GUCACGU   C   CUUCUUC       1546   GGUGAAG   CUGAUGAG   X   CGAA   AGGACGU   ACGUCCU   U   CUUCACC       1547   GGGUGAA   CUGAUGAG   X   CGAA   AAGGACG   CGUCCUU   C   UUCACCC       1549   UUGGGUG   CUGAUGAG   X   CGAA   AGAAGGA   UCCUUCU   U   CACCCAA       1550   CUUGGGU   CUGAUGAG   X   CGAA   AAGAAGG   CCUUCUU   C   ACCCAAG       1574   UGAGCUG   CUGAUGAG   X   CGAA   AUUCUCU   AGAGAAU   C   CAGCUCA       1580   UGUUUAU   CUGAUGAG   X   CGAA   AGCUGGA   UCCAGCU   C   AUAAACA       1583   UGGUGUU   CUGAUGAG   X   CGAA   AUGAGCU   AGCUCAU   A   AACACCA       1607   UCCUGUU   CUGAUGAG   X   CGAA   AUGUGCC   GGCACAU   C   AACAGGA       1636   GUUGAGG   CUGAUGAG   X   CGAA   AUCCAUG   AAUGAAU   C   CCUCAAC       1640   CGGUGUU   CUGAUGAG   X   CGAA   AGGGAUU   AAUCCCU   C   AACACCG       1651   GGCAAAG   CUGAUGAG   X   CGAA   ACCCGGU   ACCGGGU   U   CUUUGCC       1652   CGGCAAA   CUGAUGAG   X   CGAA   AACCCGG   CCGGGUU   C   UUUGCCG       1654   UGCGGCA   CUGAUGAG   X   CGAA   AGAACCC   GGGUUCU   U   UGCCGCA       1655   GUGCGGC   CUGAUGAG   X   CGAA   AAGAACC   GGUUCUU   U   GCCGCAC       1666   UGCGUAG   CUGAUGAG   X   CGAA   ACAGUGC   GCACUGU   U   CUACGCA       1667   GUGCGUA   CUGAUGAG   X   CGAA   AACAGUG   CACUGUU   C   UACGCAC       1669   GUGUGCG   CUGAUGAG   X   CGAA   AGAACAG   CUGUUCU   A   CGCACAC       1681   CGAGUUG   CUGAUGAG   X   CGAA   ACUUGUG   CACAAGU   U   CAACUCG       1682   ACGAGUU   CUGAUGAG   X   CGAA   AACUUGU   ACAAGUU   C   AACUCGU       1687   UCCGGAC   CUGAUGAG   X   CGAA   AGUUGAA   UUCAACU   C   GUCCGGA       1690   GCAUCCG   CUGAUGAG   X   CGAA   ACGAGUU   AACUCGU   C   CGGAUGC       1723   GUCGAUG   CUGAUGAG   X   CGAA   AGCUGCA   UGCAGCU   C   CAUCGAC       1764   GGCUCGG   CUGAUGAG   X   CGAA   AUAGGUG   CACCUAU   A   CCGAGCC       1773   AGGUCCC   CUGAUGAG   X   CGAA   AGGCUCG   CGAGCCU   A   GGGACCU       1785   GGCCUCU   CUGAUGAG   X   CGAA   AUCCAGG   CCUGGAU   C   AGAGGCC       1794   CAGCAGU   CUGAUGAG   X   CGAA   AGGCCUC   GAGGCCU   U   ACUGCUG       1861   GAAACAG   CUGAUGAG   X   CGAA   ACACUCG   CCAGUGU   A   CUGUUUC       1866   GGGGUGA   CUGAUGAG   X   CGAA   ACAGUAC   GUACUGU   U   UCACCCC       1867   UGGUGUG   CUGAUGAG   X   CGAA   AACAGUA   UACUGUU   U   CACCCCA       1868   UUGGGGU   CUGAUGAG   X   CGAA   AAACAGU   ACUGUUU   C   ACCCCAA       1955   UGUUGAG   CUGAUGAG   X   CGAA   AGCAGCA   UGCUGCU   U   CUCAACA       1956   UUGUUGA   CUGAUGAG   X   CGAA   AAGCAGC   GCUGCUU   C   UCAACAA       1958   UGUUGUU   CUGAUGAG   X   CGAA   AGAAGCA   UGCUUCU   C   AACAACA       2020   CUUGGUG   CUGAUGAG   X   CGAA   ACCCAGU   ACUGGGU   U   CACCAAG       2021   UCUUGGU   CUGAUGAG   X   CGAA   AACCCAG   CUGGGUU   C   ACCAAGA       2094   CGAAAGC   CUGAUGAG   X   CGAA   AUCCGUG   CACGGAU   U   GCUUUCG       2098   CUUCCGA   CUGAUGAG   X   CGAA   AGCAAUC   GAUUGCU   U   UCGGAAG       2099   GCUUCCG   CUGAUGAG   X   CGAA   AAGCAAU   AUUGCUU   U   CGGAAGC       2100   UGCUUCC   CUGAUGAG   X   CGAA   AAAGCAA   UUGCUUU   C   GGAAGCA       2157   AUACACC   CUGAUGAG   X   CGAA   AGGUGUU   AACACCU   A   GGUGUAU       2163   UCAACUA   CUGAUGAG   X   CGAA   ACACCUA   UAGGUGU   A   UAGUUGA       2165   AGUCAAC   CUGAUGAG   X   CGAA   AUACACC   GGUGUAU   A   GUUGACU       2168   GGUAGUC   CUGAUGAG   X   CGAA   ACUAUAC   GUAUAGU   U   GACUACC       2173   GUAUGGG   CUGAUGAG   X   CGAA   AGUCAAC   GUUGACU   A   CCCAUAC       2179   GAGCCUG   CUGAUGAG   X   CGAA   AUGGGUA   UACCCAU   A   CAGGCUC       2186   AGUGCCA   CUGAUGAG   X   CGAA   AGCCUGU   ACAGGCU   C   UGGCACU       2194   GCAGGGG   CUGAUGAG   X   CGAA   AGUGCCA   UGGCACU   A   CCCCUGC       2207   UAAAGUU   CUGAUGAG   X   CGAA   ACAGUGC   GCACUGU   C   AACUUUA       2212   GAUGGUA   CUGAUGAG   X   CGAA   AGUUGAC   GUCAACU   U   UACCAUC       2213   AGAUGGU   CUGAUGAG   X   CGAA   AAGUUGA   UCAACUU   U   ACCAUCU       2214   AAGAUGG   CUGAUGAG   X   CGAA   AAAGUUG   CAACUUU   A   CCAUCUU       2222   UAACCUU   CUGAUGAG   X   CGAA   AAGAUGG   CCAUCUU   U   AAGGUUA       2223   CUAACCU   CUGAUGAG   X   CGAA   AAAGAUG   CAUCUUU   A   AGGUUAG       2228   ACAUCCU   CUGAUGAG   X   CGAA   ACCUUAA   UUAAGGU   U   AGGAUGU       2229   UACAUCC   CUGAUGAG   X   CGAA   AACCUUA   UAAGGUU   A   GGAUGUA       2236   CCCCACA   CUGAUGAG   X   CGAA   ACAUCCU   AGGAUGU   A   UGUGGGG       2283   UCUCCUC   CUGAUGAG   X   CGAA   AGUCCAG   CUGGACU   C   GAGGAGA       2366   AACAGGG   CUGAUGAG   X   CGAA   AGUGUCU   AGACACU   U   CCCUGUU       2367   GAACAGG   CUGAUGAG   X   CGAA   AAGUGUC   GACACUU   C   CCUGUUC       2373   GUGAAGG   CUGAUGAG   X   CGAA   ACAGGGA   UCCCUGU   U   CCUUCAC       2374   GGUGAAG   CUGAUGAG   X   CGAA   AACAGGG   CCCUGUU   C   CUUCACC       2377   GGUGGUG   CUGAUGAG   X   CGAA   AGGAACA   UGUUCCU   U   CACCACC       2378   GGGUGGU   CUGAUGAG   X   CGAA   AAGGAAC   GUUCCUU   C   ACCACCC       2387   GAGCCGG   CUGAUGAG   X   CGAA   AGGGUGG   CCACCCU   A   CCGGCUC       2394   GUGGACA   CUGAUGAG   X   CGAA   AGCCGGU   ACCGGCU   C   UGUCCAC       2398   ACCAGUG   CUGAUGAG   X   CGAA   ACAGAGC   GCUCUGU   C   CACUGGU       2406   UGGAUCA   CUGAUGAG   X   CGAA   ACCAGUG   CACUGGU   U   UGAUCCA       2407   GUGGAUC   CUGAUGAG   X   CGAA   AACCAGU   ACUGGUU   U   GAUCCAC       2411   GGAGGUG   CUGAUGAG   X   CGAA   AUCAAAC   GUUUGAU   C   CACCUCC       2443   GUACAGG   CUGAUGAG   X   CGAA   ACUCCAC   GUGCAGU   A   CCUGUAC       2449   UAUACCG   CUGAUGAG   X   CGAA   ACAGGUA   UACCUGU   A   CGGUAUA       2454   GACCCUA   CUGAUGAG   X   CGAA   ACCGUAC   GUACGGU   A   UAGGGUC       2456   CUGACCC   CUGAUGAG   X   CGAA   AUACCGU   ACGGUAU   A   GGGUCAG       2461   AACCGCU   CUGAUGAG   X   CGAA   ACCCUAU   AUAGGGU   C   AGCGGUU       2468   AGGAGAC   CUGAUGAG   X   CGAA   ACCGCUG   CAGCGGU   U   GUCUCCU       2471   CAAAGGA   CUGAUGAG   X   CGAA   ACAACCG   CGGUUGU   C   UCCUUUG       2473   CACAAAG   CUGAUGAG   X   CGAA   AGACAAC   GUUGUCU   C   CUUUGUG       2476   GAUCACA   CUGAUGAG   X   CGAA   AGGAGAC   GUCUCCU   U   UGUGAUC       2477   UGAUCAC   CUGAUGAG   X   CGAA   AAGGAGA   UCUCCUU   U   GUGAUCA       2483   CCCAUUU   CUGAUGAG   X   CGAA   AUCACAA   UUGUGAU   C   AAAUGGG       2494   CACGAUA   CUGAUGAG   X   CGAA   ACUCCCA   UGGGAGU   A   UAUCGUG       2496   AACACGA   CUGAUGAG   X   CGAA   AUACUCC   GGAGUAU   A   UCGUGUU       2498   GCAACAC   CUGAUGAG   X   CGAA   AUAUACU   AGUAUAU   C   GUGUUGC       2503   GAAAAGC   CUGAUGAG   X   CGAA   ACACGAU   AUCGUGU   U   GCUUUUC       2507   GAAGGAA   CUGAUGAG   X   CGAA   AGCAACA   UGUUGCU   U   UUCCUUC       2508   AGAAGGA   CUGAUGAG   X   CGAA   AAGCAAC   GUUGCUU   U   UCCUUCU       2509   GAGAAGG   CUGAUGAG   X   CGAA   AAAGCAA   UUGCUUU   U   CCUUCUC       2510   GGAGAAG   CUGAUGAG   X   CGAA   AAAAGCA   UGCUUUU   C   CUUCUCC       2513   CCAGGAG   CUGAUGAG   X   CGAA   AGGAAAA   UUUUCCU   U   CUCCUGG       2514   GCCAGGA   CUGAUGAG   X   CGAA   AAGGAAA   UUUCCUU   C   UCCUGGC       2516   CCGCCAG   CUGAUGAG   X   CGAA   AGAAGGA   UCCUUCU   C   CUGGCGG       2545   CAUCCAC   CUGAUGAG   X   CGAA   AGCAGGC   GCCUGCU   U   GUGGAUG       2564   CCUGGGC   CUGAUGAG   X   CGAA   AUCAGCA   UGCUGAU   A   GCCCAGG       2614   GGCCAGG   CUGAUGAG   X   CGAA   ACGCCGC   GCGGCGU   C   CCUGGCC       2636   AGGAGAG   CUGAUGAG   X   CGAA   AUGCCAU   AUGGCAU   U   CUCUCCU       2637   AAGGAGA   CUGAUGAG   X   CGAA   AAUGCCA   UGGCAUU   C   UCUCCUU       2639   GGAAGGA   CUGAUGAG   X   CGAA   AGAAUGC   GCAUUCU   C   UCCUUCC       2641   AAGGAAG   CUGAUGAG   X   CGAA   AGAGAAU   AUUCUCU   C   CUUCCUU       2644   CACAAGG   CUGAUGAG   X   CGAA   AGGAGAG   CUCUCCU   U   CCUUGUG       2645   ACACAAG   CUGAUGAG   X   CGAA   AAGGAGA   UCUCCUU   C   CUUGUGU       2648   AAAACAC   CUGAUGAG   X   CGAA   AGGAAGG   CCUUCCU   U   GUGUUUU       2653   ACAGAAA   CUGAUGAG   X   CGAA   ACACAAG   CUUGUGU   U   UUUCUGU       2654   CACAGAA   CUGAUGAG   X   CGAA   AACACAA   UUGUGUU   U   UUCUGUG       2655   GCACAGA   CUGAUGAG   X   CGAA   AAACACA   UGUGUUU   U   UCUGUGC       2656   GGCACAG   CUGAUGAG   X   CGAA   AAAACAC   GUGUUUU   U   CUGUGCC       2657   CGGCACA   CUGAUGAG   X   CGAA   AAAAACA   UGUUUUU   C   UGUGCCG       2732   GGAGCAG   CUGAUGAG   X   CGAA   AGCAGCG   CGCUGCU   C   CUGCUCC       2749   UGGUGGU   CUGAUGAG   X   CGAA   ACGCCAG   CUGGCGU   U   ACCACCA       2750   GUGGUGG   CUGAUGAG   X   CGAA   AACGCCA   UGGCGUU   A   CCACCAC       2791   UCCACAC   CUGAUGAG   X   CGAA   AUGCAGC   GCUGCAU   C   GUGUGGA       2807   CUACAAA   CUGAUGAG   X   CGAA   ACCACCC   GGGUGGU   U   UUUGUAG       2808   CCUACAA   CUGAUGAG   X   CGAA   AACCACC   GGUGGUU   U   UUGUAGG       2809   ACCUACA   CUGAUGAG   X   CGAA   AAACCAC   GUGGUUU   U   UGUAGGU       2810   GACCUAC   CUGAUGAG   X   CGAA   AAAACCA   UGGUUUU   U   GUAGGUC       2813   UUAGACC   CUGAUGAG   X   CGAA   ACAAAAA   UUUUUGU   A   GGUCUAA       2817   AGUAUUA   CUGAUGAG   X   CGAA   ACCUACA   UGUAGGU   C   UAAUACU       2819   AGAGUAU   CUGAUGAG   X   CGAA   AGACCUA   UAGGUCU   A   AUACUCU       2822   UCAAGAG   CUGAUGAG   X   CGAA   AUUAGAC   GUCUAAU   A   CUCUUGA       2825   AGGUCAA   CUGAUGAG   X   CGAA   AGUAUUA   UAAUACU   C   UUGACCU       2827   CAAGGUC   CUGAUGAG   X   CGAA   AGAGUAU   AUACUCU   U   GACCUUG       2833   UGGUGAC   CUGAUGAG   X   CGAA   AGGUCAA   UUGACCU   U   GUCACCA       2836   GUGUGGU   CUGAUGAG   X   CGAA   ACAAGGU   ACCUUGU   C   ACCACAC       2845   CACUUUG   CUGAUGAG   X   CGAA   AGUGUGG   CCACACU   A   CAAAGUG       2854   GGCGAGG   CUGAUGAG   X   CGAA   ACACUUU   AAAGUGU   U   CCUCGCC       2855   UGGCGAG   CUGAUGAG   X   CGAA   AACACUU   AAGUGUU   C   CUCGCCA       2858   GCCUGGC   CUGAUGAG   X   CGAA   AGGAACA   UGUUCCU   C   GCCAGGC       2867   ACCAUAU   CUGAUGAG   X   CGAA   AGCCUGG   CCAGGCU   C   AUAUGGU       2870   ACCACCA   CUGAUGAG   X   CGAA   AUGAGCC   GGCUCAU   A   UGGUGGU       2889   CUGGUGA   CUGAUGAG   X   CGAA   AAAGUAU   AUACUUU   A   UCACCAG       2891   CCCUGGU   CUGAUGAG   X   CGAA   AUAAAGU   ACUUUAU   C   ACCAGGG       2993   CAAAGAU   CUGAUGAG   X   CGAA   AGCUCUG   CAGAGCU   A   AUCUUUG       2996   UGUCAAA   CUGAUGAG   X   CGAA   AUUAGCU   AGCUAAU   C   UUUGACA       2998   AAUGUCA   CUGAUGAG   X   CGAA   AGAUUAG   CUAAUCU   U   UGACAUU       2999   UAAUGUC   CUGAUGAG   X   CGAA   AAGAUUA   UAAUCUU   U   GACAUUA       3005   GUUUGGU   CUGAUGAG   X   CGAA   AUGUCAA   UUGACAU   U   ACCAAAC       3006   AGUUUGG   CUGAUGAG   X   CGAA   AAUGUCA   UGACAUU   A   CCAAACU       3014   CGAGCAG   CUGAUGAG   X   CGAA   AGUUUGG   CCAAACU   C   CUGCUCG       3020   GAAUGGC   CUGAUGAG   X   CGAA   AGCAGGA   UCCUGCU   C   GCCAUUC       3026   GACCGAG   CUGAUGAG   X   CGAA   AUGGCGA   UCGCCAU   U   CUCGGUC       3027   GGACCGA   CUGAUGAG   X   CGAA   AAUGGCG   CGCCAUU   C   UCGGUCC       3029   GCGGACC   CUGAUGAG   X   CGAA   AGAAUGG   CCAUUCU   C   GGUCCGC       3033   AUGAGCG   CUGAUGAG   X   CGAA   ACCGAGA   UCUCGGU   C   CGCUCAU       3038   GCACCAU   CUGAUGAG   X   CGAA   AGCGGAC   GUCCGCU   C   AUGGUGC       3047   CAGCCUG   CUGAUGAG   X   CGAA   AGCACCA   UGGUGCU   C   CAGGCUG       3073   UACAAAG   CUGAUGAG   X   CGAA   ACGGCAU   AUGCCGU   A   CUUUGUA       3076   GCGUACA   CUGAUGAG   X   CGAA   AGUACGG   CCGUACU   U   UGUACGC       3077   CGCGUAC   CUGAUGAG   X   CGAA   AAGUACG   CGUACUU   U   GUACGCG       3080   GAGCGCG   CUGAUGAG   X   CGAA   ACAAAGU   ACUUUGU   A   CGCGCUC       3087   AGCCCCU   CUGAUGAG   X   CGAA   AGCGCGU   ACGCGCU   C   AGGGGCU       3095   CACGAAU   CUGAUGAG   X   CGAA   AGCCCCU   AGGGGCU   U   AUUCGUG       3096   GCACGAA   CUGAUGAG   X   CGAA   AAGCCCC   GGGGCUU   A   UUCGUGC       3098   AUGCACG   CUGAUGAG   X   CGAA   AUAAGCC   GGCUUAU   U   CGUGCAU       3099   CAUGCAC   CUGAUGAG   X   CGAA   AAUAAGC   GCUUAUU   C   GUGCAUG       3112   CCGCACC   CUGAUGAG   X   CGAA   ACAUGCA   UGCAUGU   U   GGUGCGG       3125   CUCCGGC   CUGAUGAG   X   CGAA   ACUUUCC   GGAAAGU   A   GCCGGAG       3180   ACGUACG   CUGAUGAG   X   CGAA   ACCUGUC   GACAGGU   A   CGUACGU       3184   AUAGACG   CUGAUGAG   X   CGAA   ACGUACC   GGUACGU   A   CGUCUAU       3188   GGUCAUA   CUGAUGAG   X   CGAA   ACGUACG   CGUACGU   C   UAUGACC       3190   AUGGUCA   CUGAUGAG   X   CGAA   AGACGUA   UACGUCU   A   UGACCAU       3198   GGGGUAA   CUGAUGAG   X   CGAA   AUGGUCA   UGACCAU   C   UUACCCC       3200   GCGGGGU   CUGAUGAG   X   CGAA   AGAUGGU   ACCAUCU   U   ACCCCGC       3201   AGCGGGG   CUGAUGAG   X   CGAA   AAGAUGG   CCAUCUU   A   CCCCGCU       3254   CGGGCUC   CUGAUGAG   X   CGAA   ACUGCCA   UGGCAGU   A   GAGCCCG       3269   UGUCAGA   CUGAUGAG   X   CGAA   AAGACGA   UCGUCUU   C   UCUGACA       3271   CAUGUCA   CUGAUGAG   X   CGAA   AGAAGAC   GUCUUCU   C   UGACAUG       3374   GUCCCAG   CUGAUGAG   X   CGAA   AGUAUCU   AGAUACU   U   CUGGGAC       3375   GGUCCCA   CUGAUGAG   X   CGAA   AAGUAUC   GAUACUU   C   UGGGACC       3390   UCAAUGC   CUGAUGAG   X   CGAA   AUCGGCC   GGCCGAU   A   GCAUUGA       3395   GCCCUUC   CUGAUGAG   X   CGAA   AUGCUAU   AUAGCAU   U   GAAGGGC       3436   UUGGGCG   CUGAUGAG   X   CGAA   AGGCCGU   ACGGCCU   A   CGCCCAA       3458   AACCAAG   CUGAUGAG   X   CGAA   AGGCCCC   GGGGCCU   A   CUUGGUU       3461   UGCAACC   CUGAUGAG   X   CGAA   AGUAGGC   GCCUACU   U   GGUUGCA       3465   ACAAUGC   CUGAUGAG   X   CGAA   ACCAAGU   ACUUGGU   U   GCAUUGU       3470   UAGUAAC   CUGAUGAG   X   CGAA   AUGCAAC   GUUGCAU   U   GUUACUA       3473   GGCUAGU   CUGAUGAG   X   CGAA   ACAAUGC   GCAUUGU   U   ACUAGCC       3474   AGGCUAG   CUGAUGAG   X   CGAA   AACAAUG   CAUUGUU   A   CUAGCCU       3477   GUGAGGC   CUGAUGAG   X   CGAA   AGUAACA   UGUUACU   A   GCCUCAC       3506   CCCCUUC   CUGAUGAG   X   CGAA   ACCUGGU   ACCAGGU   C   GAAGGGG       3544   CAGGAAA   CUGAUGAG   X   CGAA   AUUGUGU   ACACAAU   C   UUUCCUG       3546   GCCAGGA   CUGAUGAG   X   CGAA   AGAUUGU   ACAAUCU   U   UCCUGGC       3547   CGCCAGG   CUGAUGAG   X   CGAA   AAGAUUG   CAAUCUU   U   CCUGGCG       3548   UCGCCAG   CUGAUGAG   X   CGAA   AAAGAUU   AAUCUUU   C   CUGGCGA       3563   CACCAUU   CUGAUGAG   X   CGAA   ACGCAGG   CCUGCGU   U   AAUGGUG       3564   ACACCAU   CUGAUGAG   X   CGAA   AACGCAG   CUGCGUU   A   AUGGUGU       3584   CGUGGAA   CUGAUGAG   X   CGAA   ACGGUCC   GGACCGU   C   UUCCACG       3586   GCCGUGG   CUGAUGAG   X   CGAA   AGACGGU   ACCGUCU   U   CCACGGC       3587   CGCCGUG   CUGAUGAG   X   CGAA   AAGACGG   CCGUCUU   C   CACGGCG       3632   UUUGGGU   CUGAUGAG   X   CGAA   AUUGGGC   GCCCAAU   C   ACCCAAA       3643   AUUAGUG   CUGAUGAG   X   CGAA   ACAUUUG   CAAAUGU   A   CACUAAU       3648   UCUACAU   CUGAUGAG   X   CGAA   AGUGUAC   GUACACU   A   AUGUAGA       3653   CUUGGUC   CUGAUGAG   X   CGAA   ACAUUAG   CUAAUGU   A   GACCAAG       3665   AGCCGAC   CUGAUGAG   X   CGAA   AGGUCUU   AAGACCU   C   GUCGGCU       3668   GCCAGCC   CUGAUGAG   X   CGAA   ACGAGGU   ACCUCGU   C   GGCUGGC       3720   UCCGAGC   CUGAUGAG   X   CGAA   ACCGCAG   CUGCGGU   A   GCUCGGA       3758   CCGGAAU   CUGAUGAG   X   CGAA   ACGUCAG   CUGACGU   C   AUUCCGG       3815   AAUAGGA   CUGAUGAG   X   CGAA   ACGGGUC   GACCCGU   C   UCCUAUU       3817   CAAAUAG   CUGAUGAG   X   CGAA   AGACGGG   CCCGUCU   C   CUAUUUG       3820   CUUCAAA   CUGAUGAG   X   CGAA   AGGAGAC   GUCUCCU   A   UUUGAAG       3822   CCCUUCA   CUGAUGAG   X   CGAA   AUAGGAG   CUCCUAU   U   UGAAGGG       3823   GCCCUUC   CUGAUGAG   X   CGAA   AAUAGGA   UCCUAUU   U   GAAGGGC       3832   ACCCGAA   CUGAUGAG   X   CGAA   AGCCCUU   AAGGGCU   C   UUCGGGU       3834   CCACCCG   CUGAUGAG   X   CGAA   AGAGCCC   GGGCUCU   U   CGGGUGG       3925   GGGUAUG   CUGAUGAG   X   CGAA   AGUCCAC   GUGGACU   U   CAUACCC       3926   CGGGUAU   CUGAUGAG   X   CGAA   AAGUCCA   UGGACUU   C   AUACCCG       3929   CAACGGG   CUGAUGAG   X   CGAA   AUGAAGU   ACUUCAU   A   CCCGUUG       3935   UAGACUC   CUGAUGAG   X   CGAA   ACGGGUA   UACCCGU   U   GAGUCUA       3940   UUCCAUA   CUGAUGAG   X   CGAA   ACUCAAC   GUUGAGU   C   UAUGGAA       3942   GUUUCCA   CUGAUGAG   X   CGAA   AGACUCA   UGAGUCU   A   UGGAAAC       3951   CGCAUAG   CUGAUGAG   X   CGAA   AGUUUCC   GGAAACU   A   CUAUGCG       3954   GACCGCA   CUGAUGAG   X   CGAA   AGUAGUU   AACUACU   A   UGCGGUC       3961   GACCGGG   CUGAUGAG   X   CGAA   ACCGCAU   AUGCGGU   C   CCCGGUC       3968   CCGUGAA   CUGAUGAG   X   CGAA   ACCGGGG   CCCCGGU   C   UUCACGG       3970   GUCCGUG   CUGAUGAG   X   CGAA   AGACCGG   CCGGUCU   U   CACGGAC       3971   UGUCCGU   CUGAUGAG   X   CGAA   AAGACCG   CGGUCUU   C   ACGGACA       3982   GGGAGAU   CUGAUGAG   X   CGAA   AGUUGUC   GACAACU   C   AUCUCCC       3985   CGGGGGA   CUGAUGAG   X   CGAA   AUGAGUU   AACUCAU   C   UCCCCCG       3987   GCCGGGG   CUGAUGAG   X   CGAA   AGAUGAG   CUCAUCU   C   CCCCGGC       3998   UCUGCGG   CUGAUGAG   X   CGAA   ACGGCCG   CGGCCGU   A   CCGCAGA       4009   CACUUGG   CUGAUGAG   X   CGAA   AUGUCUG   CAGACAU   U   CCAAGUG       4010   CCACUUG   CUGAUGAG   X   CGAA   AAUGUCU   AGACAUU   C   CAAGUGG       4023   GCGUGUA   CUGAUGAG   X   CGAA   AUGGGCC   GGCCCAU   C   UACACGC       4025   GAGCGUG   CUGAUGAG   X   CGAA   AGAUGGG   CCCAUCU   A   CACGCUC       4032   CCAGUGG   CUGAUGAG   X   CGAA   AGCGUGU   ACACGCU   C   CCACUGG       4094   GGACGAG   CUGAUGAG   X   CGAA   ACCUUGU   ACAAGGU   A   CUCGUCC       4097   UCAGGAC   CUGAUGAG   X   CGAA   AGUACCU   AGGUACU   C   GUCCUGA       4100   GGUUCAG   CUGAUGAG   X   CGAA   ACGAGUA   UACUCGU   C   CUGAACC       4111   GGCAACA   CUGAUGAG   X   CGAA   AUGGGUU   AACCCAU   C   UGUUGCC       4126   AAAACCC   CUGAUGAG   X   CGAA   AGGUGGC   GCCACCU   U   GGGUUUU       4131   GCCCCAA   CUGAUGAG   X   CGAA   ACCCAAG   CUUGGGU   U   UUGGGGC       4132   CGCCCCA   CUGAUGAG   X   CGAA   AACCCAA   UUGGGUU   U   UGGGGCG       4133   ACGCCCC   CUGAUGAG   X   CGAA   AAACCCA   UGGGUUU   U   GGGGCGU       4141   AGACAUA   CUGAUGAG   X   CGAA   ACGCCCC   GGGGCGU   A   UAUGUCU       4143   UUAGACA   CUGAUGAG   X   CGAA   AUACGCC   GGCGUAU   A   UGUCUAA       4147   UGCCUUA   CUGAUGAG   X   CGAA   ACAUAUA   UAUAUGU   C   UAAGGCA       4149   UGUGCCU   CUGAUGAG   X   CGAA   AGACAUA   UAUGUCU   A   AGGCACA       4161   GGGUCGG   CUGAUGAG   X   CGAA   ACCAUGU   ACAUGGU   A   CCGACCC       4196   CCGUGGU   CUGAUGAG   X   CGAA   AUGGUCC   GGACCAU   U   ACCACGG       4197   CCCGUGG   CUGAUGAG   X   CGAA   AAUGGUC   GACCAUU   A   CCACGGG       4214   AGUACGU   CUGAUGAG   X   CGAA   AUGGGGG   CCCCCAU   C   ACGUACU       4219   GGUGGAG   CUGAUGAG   X   CGAA   ACGUGAU   AUCACGU   A   CUCCACC       4222   AUAGGUG   CUGAUGAG   X   CGAA   AGUACGU   ACGUACU   C   CACCUAU       4257   CCCCCAG   CUGAUGAG   X   CGAA   ACAUCCA   UGGAUGU   U   CUGGGGG       4258   GCCCCCA   CUGAUGAG   X   CGAA   AACAUCC   GGAUGUU   C   UGGGGGC       4270   GAUAUCA   CUGAUGAG   X   CGAA   AGGCGCC   GGCGCCU   A   UGAUAUC       4275   AUUAUGA   CUGAUGAG   X   CGAA   AUCAUAG   CUAUGAU   A   UCAUAAU       4277   AUAUUAU   CUGAUGAG   X   CGAA   AUAUCAU   AUGAUAU   C   AUAAUAU       4300   GUCAGUU   CUGAUGAG   X   CGAA   AGUGGCA   UGCCACU   C   AACUGAC       4309   GGUAGUC   CUGAUGAG   X   CGAA   AGUCAGU   ACUGACU   C   GACUACC       4314   AGGAUGG   CUGAUGAG   X   CGAA   AGUCGAG   CUCGACU   A   CCAUCCU       4319   UGCCCAG   CUGAUGAG   X   CGAA   AUGGUAG   CUACCAU   C   CUGGGCA       4328   CUGUGCC   CUGAUGAG   X   CGAA   AUGCCCA   UGGGCAU   C   GGCACAG       4389   GGAGGCG   CUGAUGAG   X   CGAA   AGCGGUG   CACCGCU   A   CGCCUCC       4395   GAUCCCG   CUGAUGAG   X   CGAA   AGGCGUA   UACGCCU   C   CGGGAUC       4402   GGUAACC   CUGAUGAG   X   CGAA   AUCCCGG   CCGGGAU   C   GGUUACC       4406   GCACGGU   CUGAUGAG   X   CGAA   ACCGAUC   GAUCGGU   U   ACCGUGC       4407   GGCACGG   CUGAUGAG   X   CGAA   AACCGAU   AUCGGUU   A   CCGUGCC       4427   CCUCCUC   CUGAUGAG   X   CGAA   AUAUUUG   CAAAUAU   U   GAGGAGC       4440   UUGGACA   CUGAUGAG   X   CGAA   AGCCACC   GGUGGCU   C   UGUCCAA       4465   GCCAUAG   CUGAUGAG   X   CGAA   AGGGGAU   AUCCCCU   U   CUAUGGC       4466   UGCCAUA   CUGAUGAG   X   CGAA   AAGGGGA   UCCCCUU   C   UAUGGCA       4468   CUUGCCA   CUGAUGAG   X   CGAA   AGAAGGG   CCCUUCU   A   UGGCAAG       4512   AAAAUGA   CUGAUGAG   X   CGAA   AUGCCUU   AAGGCAU   C   UCAUUUU       4514   AGAAAAU   CUGAUGAG   X   CGAA   AGAUGCC   GGCAUCU   C   AUUUUCU       4517   GGCAGAA   CUGAUGAG   X   CGAA   AUGAGAU   AUCUCAU   U   UUCUGCC       4518   UGGCAGA   CUGAUGAG   X   CGAA   AAUGAGA   UCUCAUU   U   UCUGCCA       4519   GUGGCAG   CUGAUGAG   X   CGAA   AAAUGAG   CUCAUUU   U   CUGCCAC       4520   AGUGGCA   CUGAUGAG   X   CGAA   AAAAUGA   UCAUUUU   C   UGCCACU       4550   UUGCGGC   CUGAUGAG   X   CGAA   AGCUCAU   AUGAGCU   C   GCCGCAA       4564   GAGGCCU   CUGAUGAG   X   CGAA   ACAGCUU   AAGCUGU   C   AGGCCUC       4571   UGAUUCC   CUGAUGAG   X   CGAA   AGGCCUG   CAGGCCU   C   GGAAUCA       4602   ACGUCAA   CUGAUGAG   X   CGAA   ACCCCGG   CCGGGGU   C   UUGACGU       4604   ACACGUC   CUGAUGAG   X   CGAA   AGACCCC   GGGGUCU   U   GACGUGU       4612   UAUGACG   CUGAUGAG   X   CGAA   ACACCUC   GACGUGU   C   CGUCAUA       4637   CGAUAAC   CUGAUGAG   X   CGAA   ACAUCUC   CAGAUGU   C   GUUAUCG       4640   CCACGAU   CUGAUGAG   X   CGAA   ACGACAU   AUGUCGU   U   AUCGUGG       4641   GCCACGA   CUGAUGAG   X   CGAA   AACGACA   UGUCGUU   A   UCGUGGC       4643   UUGCCAC   CUGAUGAG   X   CGAA   AUAACGA   UCGUUAU   C   GUGGCAA       4659   GUCAUUA   CUGAUGAG   X   CGAA   AGCGUCU   AGACGCU   C   UAAUGAC       4661   CCGUCAU   CUGAUGAG   X   CGAA   AGAGCGU   ACGCUCU   A   AUGACGG       4684   CGAGUCA   CUGAUGAG   X   CGAA   AGUCACC   GGUGACU   U   UGACUCG       4685   CCGAGUC   CUGAUGAG   X   CGAA   AAGUCAC   GUGACUU   U   GACUCGG       4690   GAUCACC   CUGAUGAG   X   CGAA   AGUCAAA   UUUGACU   C   GGUGAUC       4715   UCUGGGU   CUGAUGAG   X   CGAA   ACACAUG   CAUGUGU   C   ACCCAGA       4727   UGAAAUC   CUGAUGAG   X   CGAA   ACUGUCU   AGACAGU   C   GAUUUCA       4731   AAGCUGA   CUGAUGAG   X   CGAA   AUCGACU   AGUCGAU   U   UCAGCUU       4732   CAAGCUG   CUGAUGAG   X   CGAA   AAUCGAC   GUCGAUU   U   CAGCUUG       4733   CCAAGCU   CUGAUGAG   X   CGAA   AAAUCGA   UCGAUUU   C   AGCUUGG       4738   GGGAUCC   CUGAUGAG   X   CGAA   AGCUGAA   UUCAGCU   U   GGAUCCC       4743   AAGGUGG   CUGAUGAG   X   CGAA   AUCCAAG   CUUGGAU   C   CCACCUU       4750   AAUGGUA   CUGAUGAG   X   CGAA   AGGUGGG   CCCACCU   U   UACCAUU       4751   CAAUGGU   CUGAUGAG   X   CGAA   AAGGUGG   CCACCUU   U   ACCAUUG       4752   UCAAUGG   CUGAUGAG   X   CGAA   AAAGGUG   CACCUUU   A   CCAUUGA       4757   UCCUCUC   CUGAUGAG   X   CGAA   AUGGUAA   UUACCAU   U   GAGACGA       4824   CCUCCCC   CUGAUGAG   X   CGAA   ACCCCUG   CAGGGGU   A   GGGGAGG       4835   ACCUGUA   CUGAUGAG   X   CGAA   AUGCCUC   GAGGCAU   C   UACAGGU       4837   AAACCUG   CUGAUGAG   X   CGAA   AGAUGCC   GGCAUCU   A   CAGGUUU       4843   AGUCACA   CUGAUGAG   X   CGAA   ACCUGUA   UACAGGU   U   UGUGACU       4844   GAGUCAC   CUGAUGAG   X   CGAA   AACCUGU   ACAGGUU   U   GUGACUC       4851   UCUCCCG   CUGAUGAG   X   CGAA   AGUCACA   UGUGACU   C   CGGGAGA       4867   CAUGCCC   CUGAUGAG   X   CGAA   AGGGCCG   CGGCCCU   C   GGGCAUG       4876   AGAAUCG   CUGAUGAG   X   CGAA   ACAUGCC   GGCAUGU   U   CGAUUCU       4877   AAGAAUC   CUGAUGAG   X   CGAA   AACAUGC   GCAUGUU   C   GAUUCUU       4881   ACCGAAG   CUGAUGAG   X   CGAA   AUCGAAC   GUUCGAU   U   CUUCGGU       4882   GACCGAA   CUGAUGAG   X   CGAA   AAUCGAA   UUCGAUU   C   UUCGGUC       4884   AGGACCG   CUGAUGAG   X   CGAA   AGAAUCG   CGAUUCU   U   CGGUCCU       4885   CAGGACC   CUGAUGAG   X   CGAA   AAGAAUC   GAUUCUU   C   GGUCCUG       4889   CACACAG   CUGAUGAG   X   CGAA   ACCGAAG   CUUCGGU   C   CUGUGUG       4903   CGCGUCA   CUGAUGAG   X   CGAA   AGCACUC   GAGUGCU   A   UGACGCG       5011   UUCCCAG   CUGAUGAG   X   CGAA   ACUCCAG   CUGGAGU   U   CUGGGAA       5012   UUUCCCA   CUGAUGAG   X   CGAA   AACUCCA   UGGAGUU   C   UGGGAAA       5024   CUGUGAA   CUGAUGAG   X   CGAA   ACGCUUU   AAAGCGU   C   UUCACAG       5026   GCCUGUG   CUGAUGAG   X   CGAA   AGACGCU   AGCGUCU   U   CACAGGC       5027   GGCCUGU   CUGAUGAG   X   CGAA   AAGACGC   GCGUCUU   C   ACAGGCC       5036   UGUGGGU   CUGAUGAG   X   CGAA   AGGCCUG   CAGGCCU   C   ACCCACA       5045   GGGCAUC   CUGAUGAG   X   CGAA   AUGUGGG   CCCACAU   A   GAUGCCC       5056   GGACAGG   CUGAUGAG   X   CGAA   AGUGGGC   GCCCACU   U   CCUGUCC       5057   GGGACAG   CUGAUGAG   X   CGAA   AAGUGGG   CCCACUU   C   CUGUCCC       5062   GGUUUGG   CUGAUGAG   X   CGAA   ACAGGAA   UUCCUGU   C   CCAAACC       5089   GUAAGGG   CUGAUGAG   X   CGAA   AGUUGUC   GACAACU   U   CCCUUAC       5090   GGUAAGG   CUGAUGAG   X   CGAA   AAGUUGU   ACAACUU   C   CCUUACC       5094   ACCAGGU   CUGAUGAG   X   CGAA   AGGGAAG   CUUCCCU   U   ACCUGGU       5095   UACCAGG   CUGAUGAG   X   CGAA   AAGGGAA   UUCCCUU   A   CCUGGUA       5139   GGAGGUG   CUGAUGAG   X   CGAA   AGCCUGA   UCAGGCU   C   CACCUCC       5145   CACGAUG   CUGAUGAG   X   CGAA   AGGUGGA   UCCACCU   C   CAUCGUG       5149   AUCCCAC   CUGAUGAG   X   CGAA   AUGGAGG   CCUCCAU   C   GUGGGAU       5157   CACAUUU   CUGAUGAG   X   CGAA   AUCCCAC   GUGGGAU   C   AAAUGUG       5172   CGUAUGA   CUGAUGAG   X   CGAA   ACACUUC   GAAGUGU   C   UCAUACG       5174   GCCGUAU   CUGAUGAG   X   CGAA   AGACACU   AGUGUCU   C   AUACGGC       5177   UAAGCCG   CUGAUGAG   X   CGAA   AUGAGAC   GUCUCAU   A   CGGCUUA       5183   UAGGUUU   CUGAUGAG   X   CGAA   AGCCGUA   UACGGCU   U   AAACCUA       5184   GUAGGUU   CUGAUGAG   X   CGAA   AAGCCGU   ACGGCUU   A   AACCUAC       5190   UGCAGCG   CUGAUGAG   X   CGAA   AGGUUUA   UAAACCU   A   CGCUGCA       5225   CGGCUCC   CUGAUGAG   X   CGAA   AGCCUAU   AUAGGCU   A   GGAGCCG       5234   CAUUUUG   CUGAUGAG   X   CGAA   ACGGCUC   GAGCCGU   U   CAAAAUG       5235   UCAUUUU   CUGAUGAG   X   CGAA   AACGGCU   AGCCGUU   C   AAAAUGA       5246   UGAGGGU   CUGAUGAG   X   CGAA   AUCUCAU   AUGAGAU   C   ACCCUCA       5252   GAUGUGU   CUGAUGAG   X   CGAA   AGGGUGA   UCACCCU   C   ACACAUC       5259   GUUAUGG   CUGAUGAG   X   CGAA   AUGUGUG   CACACAU   C   CCAUAAC       5264   AUUUGGU   CUGAUGAG   X   CGAA   AUGGGAU   AUCCCAU   A   ACCAAAU       5272   CAUGAUG   CUGAUGAG   X   CGAA   AUUUGGU   ACCAAAU   U   CAUCAUG       5273   CCAUGAU   CUGAUGAG   X   CGAA   AAUUUGG   CCAAAUU   C   AUCAUGG       5276   AUGCCAU   CUGAUGAG   X   CGAA   AUGAAUU   AAUUCAU   C   AUGGCAU       5290   GUCGGCC   CUGAUGAG   X   CGAA   ACAUGCA   UGCAUGU   C   GGCCGAC       5349   GCGGCCA   CUGAUGAG   X   CGAA   AGCUGCA   UGCAGCU   C   UGGCCGC       5384   CCACAAU   CUGAUGAG   X   CGAA   ACCACAC   GUGUGGU   C   AUUGUGG       5387   UACCCAC   CUGAUGAG   X   CGAA   AUGACCA   UGGUCAU   U   GUGGGUA       5394   AUGAUCC   CUGAUGAG   X   CGAA   ACCCACA   UGUGGGU   A   GGAUCAU       5402   CGGACAA   CUGAUGAG   X   CGAA   AUGAUCC   GGAUCAU   U   UUGUCCG       5403   CCGGACA   CUGAUGAG   X   CGAA   AAUGAUC   GAUCAUU   U   UGUCCGG       5404   CCCGGAC   CUGAUGAG   X   CGAA   AAAUGAU   AUCAUUU   U   GUCCGGG       5407   CCUCCCG   CUGAUGAG   X   CGAA   ACAAAAU   AUUUUGU   C   CGGGAGG       5441   GGUAGAG   CUGAUGAG   X   CGAA   ACUUCCC   GGGAAGU   C   CUCUACC       5444   CCCGGUA   CUGAUGAG   X   CGAA   AGGACUG   AAGUCCU   C   UACCGGG       5446   CUCCCGG   CUGAUGAG   X   CGAA   AGAGGAC   GUCCUCU   A   CCGGGAG       5455   UUCAUCG   CUGAUGAG   X   CGAA   ACUCCCG   CGGGAGU   U   CGAUGAA       5456   UUUCAUC   CUGAUGAG   X   CGAA   AACUCCC   GGGAGUU   C   GAUGAAA       5479   GAGGUCG   CUGAUGAG   X   CGAA   AGGCGCA   UGCGCCU   C   ACACCUC       5486   UGUAAGG   CUGAUGAG   X   CGAA   AGGUGUG   CACACCU   C   CCUUACA       5490   UCGAUGU   CUGAUGAG   X   CGAA   AGGGAGG   CCUCCCU   U   ACAUCGA       5491   UUCGAUG   CUGAUGAG   X   CGAA   AAGGGAG   CUCCCUU   A   CAUCGAA       5495   CCUGUUC   CUGAUGAG   X   CGAA   AUGUAAG   CUUACAU   C   GAACAGG       5513   GCUCGGC   CUGAUGAG   X   CGAA   AGCUGCA   UGCAGCU   C   GCCGAGC       5540   GCAACCC   CUGAUGAG   X   CGAA   AGUGCCU   AGGCACU   C   GGGUUGC       5545   UUGCAGC   CUGAUGAG   X   CGAA   ACCCGAG   CUCGGGU   U   GCUGCAA       5644   GCUGAUG   CUGAUGAG   X   CGAA   AGUUCCA   UGGAACU   U   CAUCAGC       5645   CGCUGAU   CUGAUGAG   X   CGAA   AAGUUCC   GGAACUU   C   AUCAGCG       5648   UCCCGCU   CUGAUGAG   X   CGAA   AUGAAGU   ACUUCAU   C   AGCGGGA       5657   AAUACUG   CUGAUGAG   X   CGAA   AUCCCGC   GCGGGAU   A   CAGUAUU       5662   UGCUAAA   CUGAUGAG   X   CGAA   ACUGUAU   AUACAGU   A   UUUAGCA       5664   CCUGCUA   CUGAUGAG   X   CGAA   AUACUGU   ACAGUAU   U   UAGCAGG       5665   GCCUGCU   CUGAUGAG   X   CGAA   AAUACUG   CAGUAUU   U   AGCAGGC       5666   AGCCUGC   CUGAUGAG   X   CGAA   AAAUACU   AGUAUUU   A   GCAGGCU       5677   CAGAGUG   CUGAUGAG   X   CGAA   AUAAGCC   GGCUUAU   C   CACUCUG       5682   CCAGGCA   CUGAUGAG   X   CGAA   AGUGGAU   AUCCACU   C   UGCCUGG       5702   GUGAUGC   CUGAUGAG   X   CGAA   AUCGCGG   CCGCGAU   A   GCAUCAC       5707   CAUCAGU   CUGAUGAG   X   CGAA   AUGCUAU   AUAGCAU   C   ACUGAUG       5719   GGCUGUG   CUGAUGAG   X   CGAA   AUGCCAU   AUGGCAU   U   CACAGCC       5720   AGGCUGU   CUGAUGAG   X   CGAA   AAUGCCA   UGGCAUU   C   ACAGCCU       5728   GGUGAUA   CUGAUGAG   X   CGAA   AGGCUGU   ACAGCCU   C   UAUCACC       5730   CUGGUGA   CUGAUGAG   X   CGAA   AGAGGCU   AGCCUCU   A   UCACCAG       5732   GACUGGU   CUGAUGAG   X   CGAA   AUAGAGG   CCUCUAU   C   ACCAGUC       5739   GUGAGCG   CUGAUGAG   X   CGAA   ACUGGUG   CACCAGU   C   CGCUCAC       5744   GGGUGGU   CUGAUGAG   X   CGAA   AGCGGAC   GUCCGCU   C   ACCACCC       5757   AGGAGGG   CUGAUGAG   X   CGAA   AUUCUGG   CCAGAAU   A   CCCUCCU       5762   UGAACAG   CUGAUGAG   X   CGAA   AGGGUAU   AUACCCU   C   CUGUUCA       5774   CCCCUAA   CUGAUGAG   X   CGAA   AUGUUGA   UCAACAU   C   UUAGGGG       5776   UCCCCCU   CUGAUGAG   X   CGAA   AGAUGUU   AACAUCU   U   AGGGGGA       5777   AUCCCCC   CUGAUGAG   X   CGAA   AAGAUGU   ACAUCUU   A   GGGGGAU       5796   GCGAGUU   CUGAUGAG   X   CGAA   AGCAGCC   GGCUGCU   C   AACUCGC       5808   GCACUGG   CUGAUGAG   X   CGAA   AGGAGCG   CGCUCCU   C   CCAGUGC       5820   AAGGCCG   CUGAUGAG   X   CGAA   AGCAGCA   UGCUGCU   U   CGGCCUU       5885   UGUCCAC   CUGAUGAG   X   CGAA   AGCACCU   AGGUGCU   U   GUGGACA       5894   CCGCCAG   CUGAUGAG   X   CGAA   AUGUCCA   UGGACAU   U   CUGGCGG       5895   CCCGCCA   CUGAUGAG   X   CGAA   AAUGUCC   GGACAUU   C   UGGCGGG       5986   AGGGAGC   CUGAUGAG   X   CGAA   AGUUAAC   GUUAACU   U   GCUCCCU       5999   GGGAGAG   CUGAUGAG   X   CGAA   AUGGCAG   CUGCCAU   C   CUCUCCC       6002   CGGGGGA   CUGAUGAG   X   CGAA   AGGAUGG   CCAUCCU   C   UCCCCCG       6101   CGAACGC   CUGAUGAG   X   CGAA   AUCAGCC   GGCUGAU   A   GCGUUCG       6112   ACCCCGC   CUGAUGAG   X   CGAA   AAGCGAA   UUCGCUU   C   GCGGGGU       6120   ACGUGGU   CUGAUGAG   X   CGAA   ACCCCGC   GCGGGGU   A   ACCACGU       6128   UGGGGGA   CUGAUGAG   X   CGAA   ACGUGGU   ACCACGU   U   UCCCCCA       6129   GUGGGGG   CUGAUGAG   X   CGAA   AACGUGG   CCACGUU   U   CCCCCAC       6130   CGUGGGG   CUGAUGAG   X   CGAA   AAACGUG   CACGUUU   C   CCCCACG       6142   AGGCACG   CUGAUGAG   X   CGAA   AGUGCGU   ACGCACU   A   CGUGCCU       6173   UCUGAGU   CUGAUGAG   X   CGAA   ACACGUG   CACGUGU   A   ACUCAGA       6177   AGGAUCU   CUGAUGAG   X   CGAA   AGUUACA   UGUAACU   C   AGAUCCU       6182   UGGAGAG   CUGAUGAG   X   CGAA   AUCUGAG   CUCAGAU   C   CUCUCCA       6185   GGCUGGA   CUGAUGAG   X   CGAA   AGGAUCU   AGAUCCU   C   UCCAGCC       6187   GAGGCUG   CUGAUGAG   X   CGAA   AGAGGAU   AUCCUCU   C   CAGCCUC       6194   UGAUGGU   CUGAUGAG   X   CGAA   AGGCUGG   CCAGCCU   C   ACCAUCA       6200   GCUGAGU   CUGAUGAG   X   CGAA   AUGGUGA   UCACCAU   C   ACUCAGC       6204   AGCAGCU   CUGAUGAG   X   CGAA   AGUGAUG   CAUCACU   C   AGCUGCU       6221   ACUGGUG   CUGAUGAG   X   CGAA   AGCCUCU   AGAGGCU   U   CACCAGU       6222   CACUGGU   CUGAUGAG   X   CGAA   AAGCCUC   GAGGCUU   C   ACCAGUG       6233   CCUCAUU   CUGAUGAG   X   CGAA   AUCCACU   AGUGGAU   U   AAUGAGG       6234   UCCUCAU   CUGAUGAG   X   CGAA   AAUCCAC   GUGGAUU   A   AUGAGGA       6247   UGGCGUG   CUGAUGAG   X   CGAA   AGCAGUC   GACUGCU   C   CACGCCA       6259   CGAGCCG   CUGAUGAG   X   CGAA   AGCAUGG   CCAUGCU   C   CGGCUCG       6265   UAGCCAC   CUGAUGAG   X   CGAA   AGCCGGA   UCCGGCU   C   GUGGCUA       6272   CAUCCUU   CUGAUGAG   X   CGAA   AGCCACG   CGUGGCU   A   AAGGAUG       6281   AGUCCCA   CUGAUGAG   X   CGAA   ACAUCCU   AGGAUGU   U   UGGGACU       6282   CAGUCCC   CUGAUGAG   X   CGAA   AACAUCC   GGAUGUU   U   GGGACUG       6293   CCGUGCA   CUGAUGAG   X   CGAA   AUCCAGU   ACUGGAU   A   UGCACGG       6304   GUCAGUC   CUGAUGAG   X   CGAA   ACACCGU   ACGGUGU   U   GACUGAC       6313   GGUCUUG   CUGAUGAG   X   CGAA   AGUCAGU   ACUGACU   U   CAAGACC       6314   AGGUCUU   CUGAUGAG   X   CGAA   AAGUCAG   CUGACUU   C   AAGACCU       6326   UGGACUG   CUGAUGAG   X   CGAA   AGCCAGG   CCUGGCU   C   CAGUCCA       6331   GAGCUUG   CUGAUGAG   X   CGAA   ACUGGAG   CUCCAGU   C   CAAGCUC       6338   UCGGCAG   CUGAUGAG   X   CGAA   AGCUUGG   CCAAGCU   C   CUGCCGA       6349   UCCCGGC   CUGAUGAG   X   CGAA   AUUUCGG   CCGAAAU   U   GCCGGGA       6359   AGAAAGG   CUGAUGAG   X   CGAA   ACUCCCG   CGGGAGU   C   CCUUUCU       6363   GAGAAGA   CUGAUGAG   X   CGAA   AGGGACU   AGUCCCU   U   UCUUCUC       6364   UGAGAAG   CUGAUGAG   X   CGAA   AAGGGAC   GUCCCUU   U   CUUCUCA       6365   AUGAGAA   CUGAUGAG   X   CGAA   AAAGGGA   UCCCUUU   C   UUCUCAU       6367   GCAUGAG   CUGAUGAG   X   CGAA   AGAAAGG   CCUUUCU   U   CUCAUGC       6368   GGCAUGA   CUGAUGAG   X   CGAA   AAGAAAG   CUUUCUU   C   UCAUGCC       6370   UUGGCAU   CUGAUGAG   X   CGAA   AGAAGAA   UUCUUCU   C   AUGCCAA       6385   UCCCUUG   CUGAUGAG   X   CGAA   ACCCGCG   CGCGGGU   A   CAAGGGA       6395   CCCGCCA   CUGAUGAG   X   CGAA   ACUCCCU   AGGGAGU   C   UGGCGGG       6446   GUCCGGU   CUGAUGAG   X   CGAA   AUUUGUG   CACAAAU   U   ACCGGAC       6447   UGUCCGG   CUGAUGAG   X   CGAA   AAUUUGU   ACAAAUU   A   CCGGACA       6458   CGUUUUU   CUGAUGAG   X   CGAA   ACAUGUC   GACAUGU   C   AAAAACG       6468   CUCAUGG   CUGAUGAG   X   CGAA   ACCGUUU   AAACGGU   U   CCAUGAG       6469   CCUCAUG   CUGAUGAG   X   CGAA   AACCGUU   AACGGUU   C   CAUGAUG       6479   GCCCAAC   CUGAUGAG   X   CGAA   AUCCUCA   UGAGGAU   C   GUUGGGC       6482   UAGGCCC   CUGAUGAG   X   CGAA   ACGAUCC   GGAUCGU   U   GGGCCUA       6489   CAGGUUU   CUGAUGAG   X   CGAA   AGGCCCA   UGGGCCU   A   AAACCUG       6520   GAUGGGG   CUGAUGAG   X   CGAA   ACGUUCC   GGAACGU   U   CCCCAUC       6521   UGAUGGG   CUGAUGAG   X   CGAA   AACGUUC   GAACGUU   C   CCCAUCA       6527   ACGCGUU   CUGAUGAG   X   CGAA   AUGGGGA   UCCCCAU   C   AACGCGU       6535   UGUGGUG   CUGAUGAG   X   CGAA   ACGCGUU   AACGCGU   A   CACCACA       6559   CGCCGGG   CUGAUGAG   X   CGAA   AGGGUGU   ACACCCU   C   CCCGGCG       6610   CUCCACG   CUGAUGAG   X   CGAA   ACUCUUC   GAAGAGU   A   CGUGGAG       6620   CCCGCGU   CUGAUGAG   X   CGAA   AUCUCCA   UGGAGAU   U   ACGCGGG       6621   ACCCGCG   CUGAUGAG   X   CGAA   AAUCUCC   GGAGAUU   A   CGCGGGU       6654   GUGGUCA   CUGAUGAG   X   CGAA   ACCCGUC   GACGGGU   A   UGACCAC       6689   GGGCCGG   CUGAUGAG   X   CGAA   ACCUGGC   GCCAGGU   C   CCGGCCC       6781   GACCUGG   CUGAUGAG   X   CGAA   AUGUGAC   GUCACAU   U   CCAGGUC       6854   UGGAAGU   CUGAUGAG   X   CGAA   AGCACUG   CAGUGCU   C   ACUUCCA       6858   AGCAUGG   CUGAUGAG   X   CGAA   AGUGAGC   GCUCACU   U   CCAUGCU       6859   GAGCAUG   CUGAUGAG   X   CGAA   AAGUGAG   CUCACUU   C   CAUGCUC       6866   GGUCGGU   CUGAUGAG   X   CGAA   AGCAUGG   CCAUGCU   C   ACCGACC       6877   AAUGUGG   CUGAUGAG   X   CGAA   AGGGGUC   GACCCCU   C   CCACAUU       6884   CUGCUGU   CUGAUGAG   X   CGAA   AUGUGGG   CCCACAU   U   ACAGCAG       6885   UCUGCUG   CUGAUGAG   X   CGAA   AAUGUGG   CCACAUU   A   CAGCAGA       6900   CUACGUU   CUGAUGAG   X   CGAA   AGCCGUC   GACGGCU   A   AACGUAG       6945   CUAGCUG   CUGAUGAG   X   CGAA   AGAGCUG   CAGCUCU   U   CAGCUAG       6946   GCUAGCU   CUGAUGAG   X   CGAA   AAGAGCU   AGCUCUU   C   AGCUAGC       6951   AAUUGGC   CUGAUGAG   X   CGAA   AGCUGAA   UUCAGCU   A   GCCAAUU       6969   UUCAAGG   CUGAUGAG   X   CGAA   AGGCGCA   UGCGCCU   U   CCUUGAA       6970   CUUCAAG   CUGAUGAG   X   CGAA   AAGGCGC   GCGCCUU   C   CUUGAAG       6973   UGCCUUC   CUGAUGAG   X   CGAA   AGGAAGG   CCUUCCU   U   GAAGGCA       6990   UGGUGGG   CUGAUGAG   X   CGAA   AGUGCAU   AUGCACU   A   CCCACCA       7003   GUCCGGG   CUGAUGAG   X   CGAA   AGUCAUG   CAUGACU   C   CCCGGAC       7019   CCUCGAU   CUGAUGAG   X   CGAA   AGGUCAG   CUGACCU   C   AUCGAGG       7022   UGGCCUC   CUGAUGAG   X   CGAA   AUGAGGU   ACCUCAU   C   GAGGCCA       7064   CACGGGU   CUGAUGAG   X   CGAA   AUGUUUC   GAAACAU   C   ACCCGUG       7078   AUUCUCU   CUGAUGAG   X   CGAA   ACUCCAC   GUGGAGU   C   AGAGAAU       7086   ACCACCU   CUGAUGAG   X   CGAA   AUUCUCU   AGAGAAU   A   AGGUGGU       7094   CCAAAAU   CUGAUGAG   X   CGAA   ACCACCU   AGGUGGU   A   AUUUUGG       7097   AGUCCAA   CUGAUGAG   X   CGAA   AGUACCA   UGGUAAU   U   UUGGACU       7098   GAGUCCA   CUGAUGAG   X   CGAA   AAUUACC   GGUAAUU   U   UGGACUC       7099   AGAGUCC   CUGAUGAG   X   CGAA   AAAUUAC   GUAAUUU   U   GGACUCU       7105   GUCGAAA   CUGAUGAG   X   CGAA   AGUCCAA   UUGGACU   C   UUUCGAC       7107   CGGUCCA   CUGAUGAG   X   CGAA   AGAGUCC   GGACUCU   U   UCGACCC       7108   CGGGUCG   CUGAUGAG   X   CGAA   AAGAGUC   GACUCUU   U   CGACCCG       7109   GCGGGUC   CUGAUGAG   X   CGAA   AAAGAGU   ACUCUUU   C   GACCCGC       7147   UGCAACG   CUGAUGAG   X   CGAA   AUACUUC   GAAGUAU   C   CGUUGCA       7151   CUGCUGC   CUGAUGAG   X   CGAA   ACGGAUA   UAUCCGU   U   GCAGCAG       7163   UUCGCAG   CUGAUGAG   X   CGAA   AUCUCUG   CAGAGAU   C   CUGCGAA       7174   CUUCUUG   CUGAUGAG   X   CGAA   AUUUUCG   CGAAAAU   C   CAAGAAG       7183   GGGGGGG   CUGAUGAG   X   CGAA   ACUUCUU   AAGAAGU   U   CCCCCCC       7184   CGGGGGG   CUGAUGAG   X   CGAA   AACUUCU   AGAAGUU   C   CCCCCCG       7227   AACAGUG   CUGAUGAG   X   CGAA   AGGGUUG   CAACCCU   C   CACUGUU       7240   UUUCCAG   CUGAUGAG   X   CGAA   ACUCUAA   UUAGAGU   C   CUGGAAA       7308   GGUAUUG   CUGAUGAG   X   CGAA   AGGGCCC   GGGCCCU   C   CAAUACC       7313   GAGGCGG   CUGAUGAG   X   CGAA   AUUGGAG   CUCCAAU   A   CCGCCUC       7320   UUCCGUG   CUGAUGAG   X   CGAA   AGGCGGU   ACCGCCU   C   CACGGAA       7340   UCAGAAC   CUGAUGAG   X   CGAA   ACCGUCC   GGACGGU   U   GUUCUGA       7343   CUGUCAG   CUGAUGAG   X   CGAA   ACAACCG   CGGUUGU   U   CUGACAG       7344   UCUGUCA   CUGAUGAG   X   CGAA   AACAACC   GGUUGUU   C   UGACAGA       7363   GGCAGAA   CUGAUGAG   X   CGAA   ACACGGU   ACCGUGU   C   UUCUGCC       7365   AAGGCAG   CUGAUGAG   X   CGAA   AGACACG   CGUGUCU   U   CUGCCUU       7366   CAAGGCA   CUGAUGAG   X   CGAA   AAGACAC   GUGUCUU   C   UGCCUUG       7372   CUCCGCC   CUGAUGAG   X   CGAA   AGGCAGA   UCUGCCU   U   GGCGGAG       7405   CGAUCCG   CUGAUGAG   X   CGAA   AGCUGCC   GGCAGCU   C   CGGAUCG       7446   UGAUCGG   CUGAUGAG   X   CGAA   AGGGGCG   CGCCCCU   C   CCGAUCA       7452   GAGGUCU   CUGAUGAG   X   CGAA   AUCGGGA   UCCCGAU   C   AGACCUC       7459   GUCGUCA   CUGAUGAG   X   CGAA   AGGUCUG   CAGACCU   C   UGACGAC       7480   AACGUCA   CUGAUGAG   X   CGAA   AUUCUUU   AAAGAAU   C   UGACGUU       7487   ACGACUC   CUGAUGAG   X   CGAA   ACGUCAG   CUGACGU   U   GAGUCGU       7492   GGAGUAC   CUGAUGAG   X   CGAA   ACUCAAC   GUUGAGU   C   GUACUCC       7495   GGAGGAG   CUGAUGAG   X   CGAA   ACGACUC   GAGUCGU   A   CUCCUCC       7609   CCAUGUG   CUGAUGAG   X   CGAA   AGGACAU   AUGUCCU   A   CACAUGG       7631   AUGGCGU   CUGAUGAG   X   CGAA   AUCAGGG   CCCUGAU   C   ACGCCAU       7675   GUUGCUC   CUGAUGAG   X   CGAA   ACGCGUU   AACGCGU   U   GAGCAAC       7684   CAGCAGA   CUGAUGAG   X   CGAA   AGUUGCU   AGCAACU   C   UCUGCUG       7686   CGCAGCA   CUGAUGAG   X   CGAA   AGAGUUG   CAACUCU   C   UGCUGCG       7695   UUGUGGU   CUGAUGAG   X   CGAA   ACGCAGC   GCUGCGU   C   ACCACAA       7709   UGGCAUA   CUGAUGAG   X   CGAA   ACCAUGU   ACAUGGU   C   UAUGCCA       7711   UGUGGCA   CUGAUGAG   X   CGAA   AGACCAU   AUGGUCU   A   UGCCACA       7754   CAAAGGU   CUGAUGAG   X   CGAA   ACCUUCU   AGAAGGU   C   ACCUUUG       7759   UCUGUCA   CUGAUGAG   X   CGAA   AGGUGAC   GUCACCU   U   UGACAGA       7760   GUCUGUC   CUGAUGAG   X   CGAA   AAGGUGA   UCACCUU   U   GACAGAC       7802   UCUCCUU   CUGAUGAG   X   CGAA   AGCACGU   ACGUGCU   C   AAGGAGA       7825   AACUGUG   CUGAUGAG   X   CGAA   ACGCCUU   AAGGCGU   C   CACAGUU       7822   UAGCCUU   CUGAUGAG   X   CGAA   ACUGUGG   CCACAGU   U   AAGGCUA       7833   UUAGCCU   CUGAUGAG   X   CGAA   AACUGUG   CACAGUU   A   AGGCUAA       7844   CGGAUAG   CUGAUGAG   X   CGAA   AGUUUAG   CUAAACU   U   CUAUCCG       7845   ACGGAUA   CUGAUGAG   X   CGAA   AAGUUUA   UAAACUU   C   UAUCCGU       7884   UUGGCCG   CUGAUGAG   X   CGAA   AUGUGGG   CCCACAU   U   CGGCCAA       7885   UUUGGCC   CUGAUGAG   X   CGAA   AAUGUGG   CCACAUU   C   GGCCAAA       7922   GGUUCCG   CUGAUGAG   X   CGAA   ACGUCCU   AGGACGU   C   CGGAACC       7931   UGCUGGA   CUGAUGAG   X   CGAA   AGGUUCC   GGAACCU   A   UCCAGCA       7933   CUUGCUG   CUGAUGAG   X   CGAA   AUAGGUU   AACCUAU   C   CAGCAAG       7946   UGUGGUU   CUGAUGAG   X   CGAA   AUGGCCU   AGGCCAU   U   AACCACA       7947   AUGUGGU   CUGAUGAG   X   CGAA   AAUGGCC   GGCCAUU   A   ACCACAU       8000   UGGUGUC   CUGAUGAG   X   CGAA   AUUGGUG   CACCAAU   U   GACACCA       8012   UUGCCAU   CUGAUGAG   X   CGAA   AUGGUGG   CCACCAU   C   AUGGCAA       8030   CGCAGAA   CUGAUGAG   X   CGAA   ACUUCAC   GUGAAGU   U   UUCUGCG       8031   ACGCAGA   CUGAUGAG   X   CGAA   AACUUCA   UGAAGUU   U   UCUGCGU       8032   GACGCAG   CUGAUGAG   X   CGAA   AAACUUC   GAAGUUU   U   CUGCGUC       8033   GGACGCA   CUGAUGAG   X   CGAA   AAAACUU   AAGUUUU   C   UGCGUCC       8039   CCGGUUG   CUGAUGAG   X   CGAA   ACGCAGA   UCUGCGU   C   CAACCGG       8070   AUAAGGC   CUGAUGAG   X   CGAA   AGCUGGC   GCCAGCU   C   GCCUUAU       8081   CUGGGAA   CUGAUGAG   X   CGAA   ACGAUAA   UUAUCGU   A   UUCCCAG       8083   GUCUGGG   CUGAUGAG   X   CGAA   AUACGAU   AUCGUAU   U   CCCAGAC       8084   GGUCUGG   CUGAUGAG   X   CGAA   AAUACGA   UCGUAUU   C   CCAGACC       8099   AUACACG   CUGAUGAG   X   CGAA   ACUCCCA   UGGGAGU   U   CGUGUAU       8100   CAUACAC   CUGAUGAG   X   CGAA   AACUCCC   GGGAGUU   C   GUGUAUG       8105   UCUCGCA   CUGAUGAG   X   CGAA   ACACGAA   UUCGUGU   A   UGCGAGA       8121   UCGUAAA   CUGAUGAG   X   CGAA   AGCCAUU   AAUGGCU   C   UUUACGA       8123   CGUCGUA   CUGAUGAG   X   CGAA   AGAGCCA   UGGCUCU   U   UACGACG       8124   ACCUCCU   CUGAUGAG   X   CGAA   AAGAGCC   GGCUCUU   U   ACGACGU       8125   CACGUCG   CUGAUGAG   X   CGAA   AAAGAGC   GCUCUUU   A   CGACGUG       8135   GGGUGGA   CUGAUGAG   X   CGAA   ACCACGU   ACGUGGU   C   UCCACCC       8137   AAGGGUG   CUGAUGAG   X   CGAA   AGACCAC   GUGGUCU   C   CACCCUU       8144   CCUGAGG   CUGAUGAG   X   CGAA   AGGGUGG   CCACCCU   U   CCUCAGG       8145   GCCUGAG   CUGAUGAG   X   CGAA   AAGGGUG   CACCCUU   C   CUCAGGC       8148   ACGGCCU   CUGAUGAG   X   CGAA   AGGAAGG   CCUUCCU   C   AGGCCGU       8164   GUACGAG   CUGAUGAG   X   CGAA   AGCCCAU   AUGGGCU   C   CUCGUAC       8167   UCCGUAC   CUGAUGAG   X   CGAA   AGGAGCC   GGCUCCU   C   GUACGGA       8177   AGUACUG   CUGAUGAG   X   CGAA   AAUCCGU   ACGGAUU   C   CAGUACU       8185   CCCAGGA   CUGAUGAG   X   CGAA   AGUACUG   CAGUACU   C   UCCUGGG       8241   AAGCCCA   CUGAUGAG   X   CGAA   AGGGCUU   AAGCCCU   A   UGGGCUU       8248   AUACGAG   CUGAUGAG   X   CGAA   AGCCCAU   AUGGGCU   U   CUCGUAU       8249   CAUACGA   CUGAUGAG   X   CGAA   AAGCCCA   UGGGCUU   C   UCGUAUG       8251   GUCAUAC   CUGAUGAG   X   CGAA   AGAAGCC   GGCUUCU   C   GUAUGAC       8254   GGUGUCA   CUGAUGAG   X   CGAA   ACGAGAA   UUCUCGU   A   UGACACC       8269   UGAGUCA   CUGAUGAG   X   CGAA   AGCAGCG   CGCUGCU   U   UGACUCA       8270   UUGAGUC   CUGAUGAG   X   CGAA   AAGCAGC   GCUGCUU   U   GACUCAA       8275   GACUGUU   CUGAUGAG   X   CGAA   AGUCAAA   UUUGACU   C   AACAGUC       8282   UCUCAGU   CUGAUGAG   X   CGAA   ACUGUUG   CAACAGU   C   ACUGAGA       8297   CAACACG   CUGAUGAG   X   CGAA   AUGUCGC   GCGACAU   C   CGUGUUG       8303   ACUCCUC   CUGAUGAG   X   CGAA   ACACGGA   UCCGUGU   U   GAGGAGU       8311   GUAGAUU   CUGAUGAG   X   CGAA   ACUCCUC   GAGGAGU   C   AAUCUAC       8315   AUUGGUA   CUGAUGAG   X   CGAA   AUUGACU   AGUCAAU   C   UACCAAU       8317   ACAUUGG   CUGAUGAG   X   CGAA   AGAUUGA   UCAAUCU   A   CCAAUGU       8325   AAGUCAC   CUGAUGAG   X   CGAA   ACAUUGG   CCAAUGU   U   GUGACUU       8332   GGGGGCC   CUGAUGAG   X   CGAA   AGUCACA   UGUGACU   U   GGCCCCC       8400   UUUGAAU   CUGAUGAG   X   CGAA   AGUCAGG   CCUGACU   A   AUUCAAA       8403   CCUUUUG   CUGAUGAG   X   CGAA   AUUAGUC   GACUAAU   U   CAAAAGG       8404   CCCUUUU   CUGAUGAG   X   CGAA   AAUUAGU   ACUAAUU   C   AAAAGGG       8472   GUGAGGG   CUGAUGAG   X   CGAA   AUUGCCG   CGGCAAU   A   CCCUCAC       8477   AGCAUGU   CUGAUGAG   X   CGAA   AGGGUAU   AUACCCU   C   ACAUGCU       8485   UUUCAAG   CUGAUGAG   X   CGAA   AGCAUGU   ACAUGCU   A   CUUGAAA       8488   GGCUUUC   CUGAUGAG   X   CGAA   AGUAGCA   UGCUACU   U   GAAAGCC       8565   UCACAGA   CUGAUGAG   X   CGAA   AACGACA   UGUCGUU   A   UCUGUGA       8567   UUUCACA   CUGAUGAG   X   CGAA   AUAACGA   UCGUUAU   C   UGUGAAA       8606   AGACUCG   CUGAUGAG   X   CGAA   AGGCUCG   CGAGCCU   A   CGAGUCU       8612   CCGUGAA   CUGAUGAG   X   CGAA   ACUCGUA   UACGAGU   C   UUCACGG       8614   CUCCGUG   CUGAUGAG   X   CGAA   AGACUCG   CGAGUCU   U   CACGGAG       8615   CCUCCGU   CUGAUGAG   X   CGAA   AAGACUC   GAGUCUU   C   ACGGAGG       8625   CUAGUCA   CUGAUGAG   X   CGAA   AGCCUCC   GGAGGCU   A   UGACUAG       8631   GAGUACC   CUGAUGAG   X   CGAA   AGUCAUA   UAUGACU   A   GGUACUC       8635   GGCAGAG   CUGAUGAG   X   CGAA   ACCUAGU   ACUAGGU   A   CUCUGCC       8677   CAACUCC   CUGAUGAG   X   CGAA   AGUCGUA   UACGACU   U   GGAGUUG       8683   UGUUAUC   CUGAUGAG   X   CGAA   ACUCCAA   UUGGAGU   U   GAUAACA       8687   AUGAUGU   CUGAUGAG   X   CGAA   AUCAACU   AGUUGAU   A   ACAUCAU       8692   GGAGCAU   CUGAUGAG   X   CGAA   AUGUUAU   AUAACAU   C   AUGCUCC       8710   CGCGACC   CUGAUGAG   X   CGAA   ACACGUU   AACGUGU   C   GGUCGCG       8714   CGUGCGC   CUGAUGAG   X   CGAA   ACCGACA   UGUCGGU   C   GCGCACG       8743   GAGGUAG   CUGAUGAG   X   CGAA   ACACUCU   AGAGUGU   A   CUACCUC       8746   AGUGAGG   CUGAUGAG   X   CGAA   AGUACAC   GUGUACU   A   CCUCACU       8750   CACGAGU   CUGAUGAG   X   CGAA   AGGUAGU   ACUACCU   C   ACUCGUG       8754   GGAUCAC   CUGAUGAG   X   CGAA   AGUGAGG   CCUCACU   C   GUGAUCC       8760   GUGGUGG   CUGAUGAG   X   CGAA   AUCACGA   UCGUGAU   C   CCACCAC       8799   GUGUGUC   CUGAUGAG   X   CGAA   AGCUGUC   GACAGCU   A   GACACAC       8808   UUGACUG   CUGAUGAG   X   CGAA   AGUGUGU   ACACACU   C   CAGUCAA       8813   AGGAGUU   CUGAUGAG   X   CGAA   ACUGGAG   CUCCAGU   C   AACUCCU       8818   UAGCCAG   CUGAUGAG   X   CGAA   AGUUGAC   GUCAACU   C   CUGGCUA       8825   UGUUGCC   CUGAUGAG   X   CGAA   AGCCAGG   CCUGGCU   A   GGCAACA       8834   ACAUGAU   CUGAUGAG   X   CGAA   AUGUUGC   GCAACAU   C   AUCAUGU       8837   CAUACAU   CUGAUGAG   X   CGAA   AUGAUGU   ACAUCAU   C   AUGUAUG       8870   UCAUCAA   CUGAUGAG   X   CGAA   AUCAUCC   GGAUGAU   U   UUGAUGA       8872   AGUCAUC   CUGAUGAG   X   CGAA   AAAUCAU   AUGAUUU   U   GAUGACU       8884   GGAGAAG   CUGAUGAG   X   CGAA   AGUGAGU   ACUCACU   U   CUUCUCC       8885   UGGAGAA   CUGAUGAG   X   CGAA   AAGUGAG   CUCACUU   C   UUCUCCA       8887   GAUGGAG   CUGAUGAG   X   CGAA   AGAAGUG   CACUUCU   U   CUCCAUC       8888   GGAUGGA   CUGAUGAG   X   CGAA   AAGAAGU   ACUUCUU   C   UCCAUCC       8890   AAGGAUG   CUGAUGAG   X   CGAA   AGAAGAA   UUCUUCU   C   CAUCCUG       8894   CUAGAAG   CUGAUGAG   X   CGAA   AUGGAGA   UCUCCAU   C   CUUCUAG       8897   GGGCUAG   CUGAUGAG   X   CGAA   AGGAUGG   CCAUCCU   U   CUAGCCC       8898   UGGGCUA   CUGAUGAG   X   CGAA   AAGGAUG   CAUCCUU   C   UAGCCCA       8900   CCUGGGC   CUGAUGAG   X   CGAA   AGAAGGA   UCCUUCU   A   GCCCAGG       8915   CCUUUUC   CUGAUGAG   X   CGAA   AGCUGUU   AACAGCU   U   GAAAAGG       8952   AUGGAGU   CUGAUGAG   X   CGAA   ACAGGCC   GGCCUGU   U   ACUCCAU       8953   AAUGGAG   CUGAUGAG   X   CGAA   AACAGGC   GCCUGUU   A   CUCCAUU       8956   CUCAAUG   CUGAUGAG   X   CGAA   AGUAACA   UGUUACU   C   CAUUGAG       8960   GUGGCUC   CUGAUGAG   X   CGAA   AUGGAGU   ACUCCAU   U   GAGCCAC       8969   GUAGGUC   CUGAUGAG   X   CGAA   AGUGGCU   AGCCACU   U   GACCUAC       8975   UCUGAGG   CUGAUGAG   X   CGAA   AGGUCAA   UUGACCU   A   CCUCAGA       8979   AUGAUCU   CUGAUGAG   X   CGAA   AGGUAGG   CCUACCU   C   AGAUCAU       8984   GUUGAAU   CUGAUGAG   X   CGAA   AUCUGAG   CUCAGAU   C   AUUCAAC       8987   GUCGUUG   CUGAUGAG   X   CGAA   AUGAUCU   AGAUCAU   U   CAACGAC       8988   AGUCGUU   CUGAUGAG   X   CGAA   AAUGAUC   GAUCAUU   C   AACGACU       8996   GACCAUG   CUGAUGAG   X   CGAA   AGUCGUU   AACGACU   C   CAUGGUC       9003   GCGCUAA   CUGAUGAG   X   CGAA   ACCAUGG   CCAUGGU   C   UUAGCGC       9005   AUGCGCU   CUGAUGAG   X   CGAA   AGACCAU   AUGGUCU   U   AGCGCAU       9006   AAUGCGC   CUGAUGAG   X   CGAA   AAGACCA   UGGUCUU   A   GCGCAUU       9013   GAGUGAG   CUGAUGAG   X   CGAA   AUGCGCU   AGCGCAU   U   CUCACUC       9014   GGAGUGA   CUGAUGAG   X   CGAA   AAUGCGC   GCGCAUU   C   UCACUCC       9016   AUGGAGU   CUGAUGAG   X   CGAA   AGAAUGC   GCAUUCU   C   ACUCCAU       9020   AACUAUG   CUGAUGAG   X   CGAA   AGUGAGA   UCUCACU   C   CAUAGUU       9024   GAGUAAC   CUGAUGAG   X   CGAA   AUGGAGU   ACUCCAU   A   GUUACUC       9027   GGAGAGU   CUGAUGAG   X   CGAA   ACUAUGG   CCAUAGU   U   ACUCUCC       9028   UGGAGAG   CUGAUGAG   X   CGAA   AACUAUG   CAUAGUU   A   CUCUCCA       9032   ACCUGGA   CUGAUGAG   X   CGAA   AGUAACU   AGUUACU   C   UCCAGGU       9033   UCACCUG   CUGAUGAG   X   CGAA   AGAGUAA   UUACUCU   C   CAGGUGA       9044   CCCUAUU   CUGAUGAG   X   CGAA   AUCUCAC   GUGAGAU   C   AAUAGGG       9048   GCCACCC   CUGAUGAG   X   CGAA   ACUGAUC   GAUCAAU   A   GGGUGGC       9057   AGGCAUG   CUGAUGAG   X   CGAA   AGCCACC   GGUGGCU   U   CAUGCCU       9058   GAGGCAU   CUGAUGAG   X   CGAA   AAGCCAC   GUGGCUU   C   AUGCCUC       9105   CUGGCCC   CUGAUGAG   X   CGAA   AUGUCUC   GAGACAU   C   GGGCCAG       9169   GAAGAGG   CUGAUGAG   X   CGAA   ACUUGCC   GGCAAGU   A   CCUCUUC       9173   AGUUGAA   CUGAUGAG   X   CGAA   AGGUACU   AGUACCU   C   UUCAACU       9175   CCAGUUG   CUGAUGAG   X   CGAA   AGAGGUA   UACCUCU   U   CAACUGG       9176   CCCAGUU   CUGAUGAG   X   CGAA   AAGAGGU   ACCUCUU   C   AACUGGG       9188   UGGUCCU   CUGAUGAG   X   CGAA   ACUGCCC   GGGCAGU   A   AGGACCA       9200   UGAGUUU   CUGAUGAG   X   CGAA   AGCUUGG   CCAAGCU   C   AAACUCA       9206   UUGGAGU   CUGAUGAG   X   CGAA   AGUUUGA   UCAAACU   C   ACUCCAA       9210   GGGAUUG   CUGAUGAG   X   CGAA   AGUGAGU   ACUCACU   C   CAAUCCC       9215   CGGCCGG   CUGAUGAG   X   CGAA   AUUGGAG   CUCCAAU   C   CCGGCCG       9261   CCGCUGU   CUGAUGAG   X   CGAA   ACCAGCA   UGCUGGU   U   ACAGCGG       9262   CCCGCUG   CUGAUGAG   X   CGAA   AACCAGC   GCUGGUU   A   CAGCGGG       9294   CGGGCAC   CUGAUGAG   X   CGAA   AGACAGG   CCUGUCU   C   GUGCCCG       9313   CCACAUA   CUGAUGAG   X   CGAA   ACCAGCG   CGCUGGU   U   UAUGUGG       9314   ACCACAU   CUGAUGAG   X   CGAA   AACCAGC   GCUGGUU   U   AUGUGGU       9315   CACCACA   CUGAUGAG   X   CGAA   AAACCAG   CUGGUUU   A   UGUGGUG       9409   AAAAGGG   CUGAUGAG   X   CGAA   AUGGCCU   AGGCCAU   C   CCCUUUU       9414   AAAAAAA   CUGAUGAG   X   CGAA   AGGGGAU   AUCCCCU   U   UUUUUUU                  
 
     [0299] Where “X” repesents stem II region of a HH ribozyme (Hertel et al., 1992  Nucleic Acids Res.  20: 3252). The length of stem II may be 2 base-pairs.  
               TABLE VII                          HCV Hairpin (HP) Ribozyme and Target Sequence                         Pos.   Ribozyme Sequence   Substrate                                 10   CCCCCA AGAA GGGG ACCAGAGAAACA X GUACAUUACCUGGUA   CCCC CGAU UGGGGG       59   CGUGAA AGAA GUAG ACCAGAGAAACA X GUACAUUACCUGGUA   CUAC UGUC UUCACG       109   CCUGGA AGAA GCAC ACCAGAGAAACA X GUACAUUACCUGGUA   GUGC AGCC UCCAGG       209   GCAUUG AGAA GGUU ACCAGAGAAACA X GUACAUUACCUGGUA   AACC CGCU CAAUGC       290   CUAUCA AGAA GUAC ACCAGAGAAACA X GUACAUUACCUGGUA   GUAC UGCC UGAUAG       390   GUGGGC AGAA GUAG ACCAGAGAAACA X GUACAUUACCUGGUA   CUAC CGCC GCCCAC       393   CCUGUG AGAA GCGG ACCAGAGAAACA X GUACAUUACCUGGUA   CCGC CGCC CACAGG       427   CCAACG AGAA GACC ACCAGAGAAACA X GUACAUUACCUGGUA   GGUC AGAU CGUUGG       505   GGUUGC AGAA GUUC ACCAGAGAAACA X GUACAUUACCUGGUA   GAAC GGUC GCAACC       549   CCUCGG AGAA GCGA ACCAGAGAAACA X GUACAUUACCUGGUA   UCGC CGAC CCGAGG       574   UACCCA AGAA GAGC ACCAGAGAAACA X GUACAUUACCUGGUA   GCUC AGCC UGGGUA       645   GCCGGG AGAA GCGG ACCAGAGAAACA X GUACAUUACCUGGUA   CCGC GGCU CCCGGC       652   CAACUA AGAA GGGA ACCAGAGAAACA X GUACAUUACCUGGUA   UCCC GGCC UAGUUG       671   CCGGGG AGAA GUGG ACCAGAGAAACA X GUACAUUACCUGGUA   CCAC GGAC CCCCGG       726   CGGCGA AGAA GCAC ACCAGAGAAACA X GUACAUUACCUGGUA   GUGC GGCU UCGCCG       734   CAUGAG AGAA GCGA ACCAGAGAAACA X GUACAUUACCUGGUA   UCGC CGAC CUCAUG       754   CCGACG AGAA GAAU ACCAGAGAAACA X GUACAUUACCUGGUA   AUUC CGCU CGUCGG       852   AAGAGC AGAA GGGC ACCAGAGAAACA X GUACAUUACCUGGUA   GCCC GGUU GCUCUU       883   CAGGAC AGAA GGGC ACCAGAGAAACA X GUACAUUACCUGGUA   GCCC UGCU GUCCUG       886   AAACAG AGAA GCAG ACCAGAGAAACA X GUACAUUACCUGGUA   CUGC UGUC CUGUUU       891   UGGUCA AGAA GGAC ACCAGAGAAACA X GUACAUUACCUGGUA   GUCC UGUU UGACCA       905   AGCGGA AGAA GGGA ACCAGAGAAACA X GUACAUUACCUGGUA   UCCC AGCU UCCGCU       911   CUGAUA AGAA GAAG ACCAGAGAAACA X GUACAUUACCUGGUA   CUUC CGCU UAUCAG       960   AGUUGG AGAA GUCA ACCAGAGAAACA X GUACAUUACCUGGUA   UGAC UGCU CCAACU       1050   CCCAAC AGAA GGAG ACCAGAGAAACA X GUACAUUACCUGGUA   CUCC CGUU GUUGGG       1145   GAAAGC AGAA GCCC ACCAGAGAAACA X GUACAUUACCUGGUA   GGGC GGCC GCUUUC       1148   ACAGAA AGAA GCCG ACCAGAGAAACA X GUACAUUACCUGGUA   CGGC CGCU UUCUGU       1155   UGGCGG AGAA GAAA ACCAGAGAAACA X GUACAUUACCUGGUA   UUUC UGUU CCGCCA       1185   AAACGG AGAA GCAG ACCAGAGAAACA X GUACAUUACCUGGUA   CUGC GGAU CCGUUU       1190   GAGGAA AGAA GAUC ACCAGAGAAACA X GUACAUUACCUGGUA   GAUC CGUU UUCCUC       1207   GUGAAC AGAA GGGA ACCAGAGAAACA X GUACAUUACCUGGUA   UCCC AGUU GUUCAC       1331   CACUAG AGAA GUUG ACCAGAGAAACA X GUACAUUACCUGGUA   CAAC AGCC CUAGUG       1357   UGUGGG AGAA GGAG ACCAGAGAAACA X GUACAUUACCUGGUA   CUCC GGAU CCCACA       1370   AUCCAC AGAA GCUU ACCAGAGAAACA X GUACAUUACCUGGUA   AAGC UGUC GUGGAU       1562   UCUCUG AGAA GGCC ACCAGAGAAACA X GUACAUUACCUGGUA   GGCC GGCC CAGAGA       1576   UUUAUG AGAA GGAU ACCAGAGAAACA X GUACAUUACCUGGUA   AUCC AGCU CAUAAA       1596   UGUGCC AGAA GCCA ACCAGAGAAACA X GUACAUUACCUGGUA   UGGC AGCU GGCACA       1616   GUUCAG AGAA GUCC ACCAGAGAAACA X GUACAUUACCUGGUA   GGAC UGCC CUGAAC       1663   GCGUAG AGAA GUGC ACCAGAGAAACA X GUACAUUACCUGGUA   GCAC UGUU CUACGC       1692   CUGGGC AGAA GGAC ACCAGAGAAACA X GUACAUUACCUGGUA   GUCC GGAU GCCCAG       1713   AGCUGC AGAA GGCC ACCAGAGAAACA X GUACAUUACCUGGUA   GGCC AGCU GCAGCU       1719   CGAUGG AGAA GCAG ACCAGAGAAACA X GUACAUUACCUGGUA   CUGC AGCU CCAUCG       1797   AAUGCC AGAA GUAA ACCAGAGAAACA X GUACAUUACCUGGUA   UUAC UGCU GGCAUU       1863   GGGUGA AGAA GUAC ACCAGAGAAACA X GUACAUUACCUGGUA   GUAC UGUU UCACCC       1880   CACUAC AGAA GGGC ACCAGAGAAACA X GUACAUUACCUGGUA   GCCC UGUU GUAGUG       1898   GGACCG AGAA GUCG ACCAGAGAAACA X GUACAUUACCUGGUA   CGAC CGAU CGGUCC       1903   GCACCG AGAA GAUC ACCAGAGAAACA X GUACAUUACCUGGUA   GAUC GGUC CGGUGC       1943   CAGCAC AGAA GUCU ACCAGAGAAACA X GUACAUUACCUGGUA   AGAC AGAU GUGCUG       1951   UUGAGA AGAA GCAC ACCAGAGAAACA X GUACAUUACCUGGUA   GUGC UGCU UCUCAA       1969   UGUGGC AGAA GCGU ACCAGAGAAACA X GUACAUUACCUGGUA   ACGC GGCC GCCACA       2082   CCGUGG AGAA GGUC ACCAGAGAAACA X GUACAUUACCUGGUA   GACC UGCC CCACGG       2090   AAAGCA AGAA GUGG ACCAGAGAAACA X GUACAUUACCUGGUA   CCAC GGAU UGCUUU       2316   GCUCCG AGAA GUCC ACCAGAGAAACA X GUACAUUACCUGGUA   GGAC AGAU CGGAGC       2328   GCAGCG AGAA GAGC ACCAGAGAAACA X GUACAUUACCUGGUA   GCUC AGCC CGCUGC       2332   AGCAGC AGAA GGCU ACCAGAGAAACA X GUACAUUACCUGGUA   AGCC CGCU GCUGCU       2335   GACAGC AGAA GCGG ACCAGAGAAACA X GUACAUUACCUGGUA   CCGC UGCU GCUGUC       2338   GUGGAC AGAA GCAG ACCAGAGAAACA X GUACAUUACCUGGUA   CUGC UGCU GUCCAC       2341   GUCGUG AGAA GCAG ACCAGAGAAACA X GUACAUUACCUGGUA   CUGC UGUC CACGAC       2370   UGAAGG AGAA GGGA ACCAGAGAAACA X GUACAUUACCUGGUA   UCCC UGUU CCUUCA       2390   GGACAG AGAA GGUA ACCAGAGAAACA X GUACAUUACCUGGUA   UACC GGCU CUGUCC       2395   CCAGUG AGAA GAGC ACCAGAGAAACA X GUACAUUACCUGGUA   GCUC UGUC CACUGG       2465   GGAGAC AGAA GCUG ACCAGAGAAACA X GUACAUUACCUGGUA   CAGC GGUU GUCUCC       2522   GCGCGC AGAA GCCA ACCAGAGAAACA X GUACAUUACCUGGUA   UGGC GGAC GCGCGC       2541   UCCACA AGAA GGCA ACCAGAGAAACA X GUACAUUACCUGGUA   UGCC UGCU UGUGGA       2557   GCUAUC AGAA GCAU ACCAGAGAAACA X GUACAUUACCUGGUA   AUGC UGCU GAUAGC       2579   CUCUAG AGAA GCCU ACCAGAGAAACA X GUACAUUACCUGGUA   AGGC CGCC CUAGAG       2627   AAUGCC AGAA GCUC ACCAGAGAAACA X GUACAUUACCUGGUA   GAGC GGAU GGCAUU       2663   GUACCA AGAA GCAC ACCAGAGAAACA X GUACAUUACCUGGUA   GUGC CGCC UGGUAC       2725   AGGAGC AGAA GCCA ACCAGAGAAACA X GUACAUUACCUGGUA   UGGC CGCU GCUCCU       2728   AGCAGG AGAA GCGG ACCAGAGAAACA X GUACAUUACCUGGUA   CCGC UGCU CCUGCU       2734   AGCAGG AGAA GGAG ACCAGAGAAACA X GUACAUUACCUGGUA   CUCC UGCU CCUGCU       2740   AACGCC AGAA GGAG ACCAGAGAAACA X GUACAUUACCUGGUA   CUCC UGCU GGCGUU       2978   UGGGUG AGAA GCAC ACCAGAGAAACA X GUACAUUACCUGGUA   GUGC GGCC CACCCA       3016   AUGGCG AGAA GGAG ACCAGAGAAACA X GUACAUUACCUGGUA   CUCC UGCU CGCCAU       3030   UGAGCG AGAA GAGA ACCAGAGAAACA X GUACAUUACCUGGUA   UCUC GGUC CGCUCA       3034   ACCAUG AGAA GACC ACCAGAGAAACA X GUACAUUACCUGGUA   GGUC CGCU CAUGGU       3260   GAAGAC AGAA GGCU ACCAGAGAAACA X GUACAUUACCUGGUA   AGCC CGUC GUCUUC       3340   GAGACG AGAA GUCC ACCAGAGAAACA X GUACAUUACCUGGUA   GGAC UGCC CGUCUC       3344   GGCGGA AGAA GGCA ACCAGAGAAACA X GUACAUUACCUGGUA   UGCC CGUC UCCGCC       3350   CCUUCG AGAA GAGA ACCAGAGAAACA X GUACAUUACCUGGUA   UCUC CGCC CGAAGG       3383   GCUAUC AGAA GGUC ACCAGAGAAACA X GUACAUUACCUGGUA   GACC GGCC GAUAGC       3431   GGCGUA AGAA GUGA ACCAGAGAAACA X GUACAUUACCUGGUA   UCAC GGCC UACGCC       3581   GUGGAA AGAA GUCC ACCAGAGAAACA X GUACAUUACCUGGUA   GGAC CGUC UUCCAC       3597   UCUUUG AGAA GGCG ACCAGAGAAACA X GUACAUUACCUGGUA   CGCC GGCU CAAAGA       3615   CUUUUG AGAA GGCU ACCAGAGAAACA X GUACAUUACCUGGUA   AGCC GGCC CAAAAG       3669   CAUGCC AGAA GACG ACCAGAGAAACA X GUACAUUACCUGGUA   CGUC GGCU GGCAUG       3725   AUAGAG AGAA GAGC ACCAGAGAAACA X GUACAUUACCUGGUA   GCUC GGAC CUCUAU       3752   AAUGAC AGAA GCAU ACCAGAGAAACA X GUACAUUACCUGGUA   AUGC UGAC GUCAUU       3771   CACCGC AGAA GCGC ACCAGAGAAACA X GUACAUUACCUGGUA   GCGC CGAC GCGGUG       3783   UCCCCC AGAA GUCA ACCAGAGAAACA X GUACAUUACCUGGUA   UGAC GGUC GGGGGA       3799   CUGGOG AGAA GUAG ACCAGAGAAACA X GUACAUUACCUGGUA   CUAC UGUC CCCCAG       3807   AGACGG AGAA GGGG ACCAGAGAAACA X GUACAUUACCUGGUA   CCCC AGAC CCGUCU       3812   AUAGGA AGAA GGUC ACCAGAGAAACA X GUACAUUACCUGGUA   GACC CGUC UCCUAU       3847   GGGCAG AGAA GUGG ACCAGAGAAACA X GUACAUUACCUGGUA   CCAC UGCU CUGCCC       3852   CCGAAG AGAA GAGC ACCAGAGAAACA X GUACAUUACCUGGUA   GCUC UGCC CUUCGG       3887   GCACAC AGAA GCCC ACCAGAGAAACA X GUACAUUACCUGGUA   GGGC UGCU GUGUGC       3932   AGACUC AGAA GGUA ACCAGAGAAACA X GUACAUUACCUGGUA   UACC CGUU GAGUCU       3958   ACCGGG AGAA GCAU ACCAGAGAAACA X GUACAUUACCUGGUA   AUGC GGUC CCCGGU       3965   CGUGAA AGAA GGGG ACCAGAGAAACA X GUACAUUACCUGGUA   CCCC GGUC UUCACG       3992   CGGUAC AGAA GGGG ACCAGAGAAACA X GUACAUUACCUGGUA   CCCC GGCC GUACCG       4064   GUACGC AGAA GGCA ACCAGAGAAACA X GUACAUUACCUGGUA   UGCC GGCU GCGUAC       4076   CCCUUG AGAA GCGU ACCAGAGAAACA X GUACAUUACCUGGUA   ACGC AGCC CAAGGG       4112   GGCGGC AGAA GAUG ACCAGAGAAACA X GUACAUUACCUGGUA   CAUC UGUU GCCGCC       4163   GUUGGG AGAA GUAC ACCAGAGAAACA X GUACAUUACCUGGUA   GUAC CGAC CCCAAC       4244   UCCACC AGAA GCAA ACCAGAGAAACA X GUACAUUACCUGGUA   UUGC CGAC GGUGGA       4304   AGUCGA AGAA GUUG ACCAGAGAAACA X GUACAUUACCUGGUA   CAAC UGAC UCGACU       4334   GUCCAG AGAA GUGC ACCAGAGAAACA X GUACAUUACCUGGUA   GCAC AGUC CUGGAC       4355   CGCUCC AGAA GUCU ACCAGAGAAACA X GUACAUUACCUGGUA   AGAC GGCU GGAGCG       4366   ACGACG AGAA GCGC ACCAGAGAAACA X GUACAUUACCUGGUA   GCGC GGCU CGUCGU       4441   GUGUUG AGAA GAGC ACCAGAGAAACA X GUACAUUACCUGGUA   GCUC UGUC CAACAC       4621   CCGCUA AGAA GUAU ACCAGAGAAACA X GUACAUUACCUGGUA   AUAC CGAC UAGCGG       4652   UAGAGC AGAA GUUG ACCAGAGAAACA X GUACAUUACCUGGUA   CAAC AGAC GCUCUA       4724   GAAAUC AGAA GUCU ACCAGAGAAACA X GUACAUUACCUGGUA   AGAC AGUC GAUUUC       4734   GAUCCA AGAA GAAA ACCAGAGAAACA X GUACAUUACCUGGUA   UUUC AGCU UGGAUC       4861   CCCGAG AGAA GUUC ACCAGAGAAACA X GUACAUUACCUGGUA   GAAC GGCC CUCGGG       4886   ACACAG AGAA GAAG ACCAGAGAAACA X GUACAUUACCUGGUA   CUUC GGUC CUGUGU       4937   AGUCUC AGAA GGCG ACCAGAGAAACA X GUACAUUACCUGGUA   CGCC CGCU GAGACU       4988   CUGGCA AGAA GGCA ACCAGAGAAACA X GUACAUUACCUGGUA   UGCC CGUC UGCCAG       5059   GUUUGG AGAA GGAA ACCAGAGAAACA X GUACAUUACCUGGUA   UUCC UGUC CCAAAC       5179   GGUUUA AGAA GUAU ACCAGAGAAACA X GUACAUUACCUGGUA   AUAC GGCU UAAACC       5212   CUAUAC AGAA GGGG ACCAGAGAAACA X GUACAUUACCUGGUA   CCCC UGCU GUAUAG       5231   AUUUUG AGAA GCUC ACCAGAGAAACA X GUACAUUACCUGGUA   GAGC CGUU CAAAAU       5291   CAGGUC AGAA GACA ACCAGAGAAACA X GUACAUUACCUGGUA   UGUC GGCC GACCUG       5294   CUCCAG AGAA GCCG ACCAGAGAAACA X GUACAUUACCUGGUA   CGGC CGAC CUGGAG       5345   GGCCAG AGAA GCAA ACCAGAGAAACA X GUACAUUACCUGGUA   UUGC AGCU CUGGCC       5417   AACAAC AGAA GGCC ACCAGAGAAACA X GUACAUUACCUGGUA   GGCC GGCU GUUGUU       5420   GGGAAC AGAA GCCG ACCAGAGAAACA X GUACAUUACCUGGUA   CGGC UGUU GUUCCC       5509   UCGGCG AGAA GCAU ACCAGAGAAACA X GUACAUUACCUGGUA   AUGC AGCU CGCCGA       5521   UGCUUG AGAA GCUC ACCAGAGAAACA X GUACAUUACCUGGUA   GAGC AGUU CAAGCA       5576   GGGAGC AGAA GCCU ACCAGAGAAACA X GUACAUUACCUGGUA   AGGC CGCU GCUCCC       5579   CACGGG AGAA GCGG ACCAGAGAAACA X GUACAUUACCUGGUA   CCGC UGCU CCCGUG       5683   UUCCCA AGAA GAGU ACCAGAGAAACA X GUACAUUACCUGGUA   ACUC UGCC UGGGAA       5710   AAUGCC AGAA GUGA ACCAGAGAAACA X GUACAUUACCUGGUA   UCAC UGAU GGCAUU       5723   GAUAGA AGAA GUGA ACCAGAGAAACA X GUACAUUACCUGGUA   UCAC AGCC UCUAUC       5736   UGAGCG AGAA GGUG ACCAGAGAAACA X GUACAUUACCUGGUA   CACC AGUC CGCUCA       5740   GUGGUG AGAA GACU ACCAGAGAAACA X GUACAUUACCUGGUA   AGUC CGCU CACCAC       5764   AUGUUG AGAA GGAG ACCAGAGAAACA X GUACAUUACCUGGUA   CUCC UGUU CAACAU       5792   GAGUUG AGAA GCCA ACCAGAGAAACA X GUACAUUACCUGGUA   UGGC UGCU CAACUC       5816   GGCCGA AGAA GCAC ACCAGAGAAACA X GUACAUUACCUGGUA   GUGC UGCU UCGGCC       5822   CACGAA AGAA GAAG ACCAGAGAAACA X GUACAUUACCUGGUA   CUUC GGCC UUCGUG       5966   GUCCUC AGAA GAGG ACCAGAGAAACA X GUACAUUACCUGGUA   CCUC CGCC GAGGAC       6094   GCUAUC AGAA GGUU ACCAGAGAAACA X GUACAUUACCUGGUA   AACC GGCU GAUAGC       6178   GAGAGG AGAA GAGU ACCAGAGAAACA X GUACAUUACCUGGUA   ACUC AGAU CCUCUC       6189   UGGUGA AGAA GGAG ACCAGAGAAACA X GUACAUUACCUGGUA   CUCC AGCC UCACCA       6205   UUCAGC AGAA GAGU ACCAGAGAAACA X GUACAUUACCUGGUA   ACUC AGCU GCUGAA       6208   CUCUUC AGAA GCUG ACCAGAGAAACA X GUACAUUACCUGGUA   CAGC UGCU GAAGAG       6243   GCGUGG AGAA GUCC ACCAGAGAAACA X GUACAUUACCUGGUA   GGAC UGCU CCACGC       6261   GCCACG AGAA GGAG ACCAGAGAAACA X GUACAUUACCUGGUA   CUCC GGCU CGUGGC       6308   CUUGAA AGAA GUCA ACCAGAGAAACA X GUACAUUACCUGGUA   UGAC UGAC UUCAAG       6328   AGCUUG AGAA GGAG ACCAGAGAAACA X GUACAUUACCUGGUA   CUCC AGUC CAAGCU       6340   AAUUUC AGAA GGAG ACCAGAGAAACA X GUACAUUACCUGGUA   CUCC UGCC GAAAUU       6426   CACAUG AGAA GGUG ACCAGAGAAACA X GUACAUUACCUGGUA   CACC UGCC CAUGUG       6465   UCAUGG AGAA GUUU ACCAGAGAAACA X GUACAUUACCUGGUA   AAAC GGUU CCAUGA       6599   CUCUUC AGAA GCCA ACCAGAGAAACA X GUACAUUACCUGGUA   UGGC UGCU GAAGAG       6692   UUCGGG AGAA GGGA ACCAGAGAAACA X GUACAUUACCUGGUA   UCCC GGCC CCCGAA       6727   CUGUGC AGAA GCAC ACCAGAGAAACA X GUACAUUACCUGGUA   GUGC GGUU GCACAG       6753   GGAGAG AGAA GCAC ACCAGAGAAACA X GUACAUUACCUGGUA   GUGC AGAC CUCUCC       6817   CAUGGG AGAA GUGA ACCAGAGAAACA X GUACAUUACCUGGUA   UCAC AGCU CCCAUG       6839   UGCCAC AGAA GGUU ACCAGAGAAACA X GUACAUUACCUGGUA   AACC GGAU GUGGCA       6869   GGAGGG AGAA GUGA ACCAGAGAAACA X GUACAUUACCUGGUA   UCAC CGAC CCCUCC       6939   CUGAAG AGAA GGCC ACCAGAGAAACA X GUACAUUACCUGGUA   GGCC AGCU CUUCAG       7007   GUCAGC AGAA GGGG ACCAGAGAAACA X GUACAUUACCUGGUA   CCCC GGAC GCUGAC       7013   GAUGAG AGAA GCGU ACCAGAGAAACA X GUACAUUACCUGGUA   ACGC UGAC CUCAUC       7114   GCUCGA AGAA OGUC ACCAGAGAAACA X GUACAUUACCUGGUA   GACC CGCU UCGAGC       7148   UGCUGC AGAA GAUA ACCAGAGAAACA X GUACAUUACCUGGUA   UAUC CGUU GCAGCA       7214   GUUGUA AGAA GGGC ACCAGAGAAACA X GUACAUUACCUGGUA   GCCC GGAU UACAAC       7253   GACGUA AGAA GGAC ACCAGAGAAACA X GUACAUUACCUGGUA   GUCC GGAC UACGUC       7291   GUGGUA AGAA GCAA ACCAGAGAAACA X GUACAUUACCUGGUA   UUGC CGCC UACCAC       7315   CGUGGA AGAA GUAU ACCAGAGAAACA X GUACAUUACCUGGUA   AUAC CGCC UCCACG       7337   CAGAAC AGAA GUCC ACCAGAGAAACA X GUACAUUACCUGGUA   GGAC GGUU GUCCUG       7367   CGCCAA AGAA GAAG ACCAGAGAAACA X GUACAUUACCUGGUA   CUUC UGCC UUGGCG       7401   AUCCGG AGAA GCCG ACCAGAGAAACA X GUACAUUACCUGGUA   CGGC AGCU CCGGAU       7407   CCGACG AGAA GGAG ACCAGAGAAACA X GUACAUUACCUGGUA   CUCC GGAU CGUCGG       7415   GUCAAC AGAA GACG ACCAGAGAAACA X GUACAUUACCUGGUA   CGUC GGCC GUGGAC       7418   GCUGUC AGAA GCCG ACCAGAGAAACA X GUACAUUACCUGGUA   CGGC CGUU GACAGC       7439   GGGAGG AGAA GUCG ACCAGAGAAACA X GUACAUUACCUGGUA   CGAC CGCC CCUCCC       7448   GGUCUG AGAA GGAG ACCAGAGAAACA X GUACAUUACCUGGUA   CUCC CGAU CAGACC       7453   UCAGAG AGAA GAUC ACCAGAGAAACA X GUACAUUACCUGGUA   GAUC AGAC CUCUGA       7460   ACCGUC AGAA GAGG ACCAGAGAAACA X GUACAUUACCUGGUA   CCUC UGAC GACGGU       7481   CUCAAC AGAA GAUU ACCAGAGAAACA X GUACAUUACCUGGUA   AAUC UGAC GUUGAG       7535   GCUGAG AGAA GGGU ACCAGAGAAACA X GUACAUUACCUGGUA   ACCC UGAU CUCAGC       7593   UUGAGC AGAA GACG ACCAGAGAAACA X GUACAUUACCUGGUA   CGUC UGCU GCUCAA       7596   ACADUG AGAA GCAG ACCAGAGAAACA X GUACAUUACCUGGUA   CUGC UGCU CAAUGU       7627   GGCGUG AGAA GGGC ACCAGAGAAACA X GUACAUUACCUGGUA   GCCC UGAU CACGCC       7660   UUGAUG AGAA GCUU ACCAGAGAAACA X GUACAUUACCUGGUA   AAGC UGCC CAUCAA       7687   UGACGC AGAA GAGA ACCAGAGAAACA X GUACAUUACCUGGUA   UCUC UGCU GCGUCA       7764   CUUGCA AGAA GUCA ACCAGAGAAACA X GUACAUUACCUGGUA   UGAC AGAC UGCAAG       7870   GGGGGC AGAA GCUU ACCAGAGAAACA X GUACAUUACCUGGUA   AAGC UGAC GCCCCC       7956   ACACGG AGAA GAUG ACCAGAGAAACA X GUACAUUACCUGGUA   CAUC CGCU CCGUGU       7975   UCUUCC AGAA GGUC ACCAGAGAAACA X GUACAUUACCUGGUA   GACC UGCU GGAAGA       8066   AAGGCG AGAA GGCU ACCAGAGAAACA X GUACAUUACCUGGUA   AGCC AGCU CGCCUU       8087   UCCCAG AGAA GGGA ACCAGAGAAACA X GUACAUUACCUGGUA   UCCC AGAC CUGGGA       8172   ACUGGA AGAA GUAC ACCAGAGAAACA X GUACAUUACCUGGUA   GUAC GGAU UCCAGU       8262   CAAAGC AGAA GGUG ACCAGAGAAACA X GUACAUUACCUGGUA   CACC CGCU GCUUUG       8265   AGUCAA AGAA GCGG ACCAGAGAAACA X GUACAUUACCUGGUA   CCGC UGCU UUGACU       8374   AUGUAG AGAA GCUC ACCAGAGAAACA X GUACAUUACCUGGUA   GAGC GGCU CUACAU       8395   GAAUUA AGAA GGGG ACCAGAGAAACA X GUACAUUACCUGGUA   CCCC UGAC UAAUUC       8452   CUAGUC AGAA GCAC ACCAGAGAAACA X GUACAUUACCUGGUA   GUGC UGAC GACUAG       8501   UCGACA AGAA GCAG ACCAGAGAAACA X GUACAUUACCUGGUA   CUGC GGCC UGUCGA       8505   CAGCUC AGAA GGCC ACCAGAGAAACA X GUACAUUACCUGGUA   GGCC UGUC GAGCUG       8639   GGGGGG AGAA GAGU ACCAGAGAAACA X GUACAUUACCUGGUA   ACUC UGCC CCCCCC       8656   GGUUGG AGAA GGUC ACCAGAGAAACA X GUACAUUACCUGGUA   GACC CGCC CCAACC       8711   GUGCGC AGAA GACA ACCAGAGAAACA X GUACAUUACCUGGUA   UGUC GGUC GCGCAC       8911   UUUUCA AGAA GUUC ACCAGAGAAACA X GUACAUUACCUGGUA   GAAC AGCU UGAAAA       8935   CCGUAG AGAA GACA ACCAGAGAAACA X GUACAUUACCUGGUA   UGUC AGAU CUACGG       8980   UGAAUG AGAA GAGG ACCAGAGAAACA X GUACAUUACCUGGUA   CCUC AGAU CAUUCA       9082   CGCAAG AGAA GUAC ACCAGAGAAACA X GUACAUUACCUGGUA   GUAC CGCC CUUGCG       9133   CCUUGG AGAA GUAG ACCAGAGAAACA X GUACAUUACCUGGUA   CUAC UGUC CCAAGG       9218   GGACGC AGAA GGGA ACCAGAGAAACA X GUACAUUACCUGGUA   UCCC GGCC GCGUCC       9229   AAGUCC AGAA GGGA ACCAGAGAAACA X GUACAUUACCUGGUA   UCCC AGCU GGACUU       9243   CGAACC AGAA GGAC ACCAGAGAAACA X GUACAUUACCUGGUA   GUCC AGCU GGUUCG       9285   GAGACA AGAA GUGA ACCAGAGAAACA X GUACAUUACCUGGUA   UCAC AGCC UGUCUC       9289   GCACGA AGAA GGCU ACCAGAGAAACA X GUACAUUACCUGGUA   AGCC UGUC UCGUGC       9300   AGCGGG AGAA GGCA ACCAGAGAAACA X GUACAUUACCUGGUA   UGCC CGAC CCCGCU       9306   UAAACC AGAA GGGU ACCAGAGAAACA X GUACAUUACCUGGUA   ACCC CGCU GGUUUA       9358   UUGGGG AGAA GGUA ACCAGAGAAACA X GUACAUUACCUGGUA   UACC UGCU CCCCAA                  
 
     [0300] Where “X”represents stem IV region of a HP ribozyme (Berzal-Herranze et al., 1993, EMBO.J. 12,2567). The length of stem IV may be 2 base-pairs.  
               TABLE VIII                          Additional HCV Conserved Hammerhead ribozyme and target sequence                                 Nos.   Name*   Pos. †     Ribozyme   Substrate                                         1   HCV.C-48   278   UUGGUGU CUGAUGAG X CGAA ACGUUUG   CAAACGU A ACACCAA       2   HCV.C-60   290   UGUGGGC CUGAUGAG X CGAA ACGGUUG   CAACCGU C GCCCACA       3   HCV.C-175   409   AGGUUGU CUGAUGAG X CGAA ACCGCUC   GAGCGGU C ACAACCU       4   HCV.3-118   9418   AAAAAAA CUGAUGAG X CGAA AAAAAAA   UUUUUUU U UUUUUUU       5   HCV.3-145   9445   UAAGAUG CUGAUGAG X CGAA AGCCACC   GGUGGCU C CAUCUUA       6   HCV.3-149   9449   GGGCUAA CUGAUGAG X CGAA AUGGAGC   GCUCCAU C UUAGCCC       7   HCV.3-151   9451   UAGGGCU CUGAUGAG X CGAA AGAUGGA   UCCAUCU U AGCCCUA       8   HCV.3-152   9452   CUAGGGC CUGAUGAG X CGAA AAGAUGG   CCAUCUU A GCCCUAG       9   HCV.3-158   9458   CCGUGAC CUGAUGAG X CGAA AGGGCUA   UAGCCCU A GUCACGG       10   HCV.3-161   9461   UAGCCGU CUGAUGAG X CGAA ACUAGGG   CCCUAGU C ACGGCUA       11   HCV.3-168   9468   UCACAGC CUGAUGAG X CGAA AGCCGUG   CACGGCU A GCUGUGA       12   HCV.3-181   9481   GCUCACG CUGAUGAG X CGAA ACCUUUC   GAAAGGU C CGUGAGC                                                                  
 
     [0301] Where “X”represents stem II region of a HH ribozyme (Hertel et al., 1992  Nucleic Acids Res.  20: 3252). The length of stem II may be 2 base-pairs.  
     [0302] Core reference Sequence for Nos. 1-3=HPCCOPR (Acc#L38318) 1-600 bp *-Nucleotide 231 (8 nucleotide upstream of the initiator ATG) has been designated as “1” for the purpose of numbering ribozyme sites in the core protein coding region.  
     [0303] 3′-NCR Reference Sequence for Nos. 4-12=D85516 (Acc#D85516) 9301-9535 bp *-Nucleotide 9301 has been designated as “1”for the purpose of numbering ribozyme sites in the 3′NCR.  
     [0304] ↑-position number reflects the reference sequence from HPCCOPR.  
               TABLE IX                          Inhibition of HCV RNA in OST7 cells Using Multiple Ribozyme Motifs                                 Motif   RPI Number   F luc /R luc     SEM   Sequence               RPI Motif I   Irrelevant Control   0.22   0.03   auccuUGAU s GGCAUACACUAUGCGCGaugaucugcaB       RPI Motif I   18738   0.13   0.03   acacuuGAU s ggcauGcacuaugcgcgauacuaacgcB       RPI Motif I   18739   0.15   0.01   cacgauGAU s ggcauGcacuaugcgcgacucauacuaB       RPI Motif I   18740   0.15   0.01   ggcuguGAU s ggcauGcacuaugcgcgacgacacucaB       RPI Motif I   18746   0.10   0.02   cccaauGAU s ggcauGcacuaugcgcgacuacucggcB       RPI Motif I   18747   0.16   0.02   uuucguGAU s ggcauGcacuaugcgcggacccaacacB       RPI Motif I   18750   0.15   0.03   ucagguGAU s ggcauGcacuaugcgcgaguaccacaaB       RPI Motif I   18754   0.12   0.01   gcacuuGAU s ggcauGcacuaugcgcggcaagcacccB       RPI Motif II   SAC   1.10   0.32   a s u s u s c s ca cUAGuGaggcguuagccGau Acgcga B       RPI Motif II   20339   0.85   0.01   u s c s c s u s caccUGAuGaggccguuaggccGaaIgggaguB       RPI Motif II   20350   1.04   0.05   g s u s c s c s uggcUGAuGaggccguuaggccGaaIgcugcaB       RPI Motif III   Irrelevant Control   1.28   0.07   ggaaaggugugcaaCCGgaggaaacucCCUUCAAGGACAUCGUCCGGGacggcB       RPI Motif III   18704   0.37   0.07   uuccgcagaCGgaggaaacucCCUUCAAGGACGAAAGUCCGGGacuauggB       RPI Motif III   18705   0.42   0.10   ccgcagaCGgaggaaacucCCUUCAAGGACGAAAGUCCGGGacuauggB       RPI Motif III   18700   0.61   0.16   cagguaguaCGgaggaaacucCCUUCAAGGACAUCGUCCGGGacaaggB       RPI Motif III   18701   0.54   0.10   gcacggucUaGgaggaaacucCCUUCAAGGACAUCGUCCGGGgagaccB       RPI Motif III   18835   0.54   0.04   guguacucacGgaggaaacucCCUUCAAGGACAUCGUCCGGGgguucB                                                                                  
 
     [0305]               TABLE X                          Anti HCV minus strand Stabilized Ribozyme and Target Sequence                             RPI                   No.   Ribozyme Alias   Ribozyme Sequence   Target Sequence               14961   HCV.5nc-34 Rz-7 allyl stab1   g s g s u s c s ucg cUGAuGaggccguuaggccGaa Agaccgu B   ACGGUCU A CGAGACC       14962   HCV.5nc-43 Rz-7 allyl stab1   g s c s c s c s cgg cUGAuGaggccguuaggccGaa Aggucuc B   GAGACCU C CCGGGGC       14963   HCV.5nc-54 Rz-7 allyl stab1   u s g s c s u s ugc cUGAuGaggccguuaggccGaa Agugccc B   GGGCACU C GCAAGCA       14964   HCV.5nc-66 Rz-7 allyl stab1   u s g s c s c s uga cUGAuGaggccguuaggccGaa Agggugc B   GCACCCU A UCAGGCA       14965   HCV.5nc-88 Rz-7 allyl stab1   g s u s c s g s cga cUGAuGaggccguuaggccGaa Aggccuu B   AAGGCCU U UCGCGAC       14966   HCV.5nc-88b Rz-7 allyl stab1   g s u s u s g s cga cUGAuGaggccguuaggccGaa Aggccuu B   AAGGCCU U UCGCAAC       14967   HCV.5nc-107 Rz-7 allyl stab1   g s c s u s a s gcc cUGAuGaggccguuaggccGaa Aguagug B   CACUACU C GGCUAGC       14968   HCV.5nc-162 Rz-7 allyl stab1   u s u s u s c s uug cUGAuGaggccguuaggccGaa Aucaacc B   GGUUGAU C CAAGAAA       14969   HCV.5nc-162b Rz-7 allyl stab1   u s u s u s c s uug cUGAuGaggccguuaggccGaa Auaaacc B   GGUUUAU C CAAGAAA       14970   HCV.5nc-192 Rz-7 allyl stab1   u s a s c s a s ccg cUGAuGaggccguuaggccGaa Aauugcc B   GGCAAUU C CGGUGUA       14971   HCV.5nc-199 Rz-7 allyl stab1   c s g s g s u s gag cUGAuGaggccguuaggccGaa Acaccgg B   CCGGUGU A CUCACCG       14972   HCV.5nc-202 Rz-7 allyl stab1   a s a s c s c s ggu cUGAuGaggccguuaggccGaa Aguacac B   GUGUACU C ACCGGUU       14973   HCV.5nc-222 Rz-7 allyl stab1   a s g s a s g s cca cUGAuGaggccguuaggccGaa Agugguc B   GACCACU A UGGCUCU       14974   HCV.5nc-265 Rz-7 allyl stab1   u s u s a s g s uau cUGAuGaggccguuaggccGaa Agugucg B   CGACACU C AUACUAA       14975   HCV.5nc-33 CHz-7 allyl stab1   g s u s c s u s cgu cUGAuGaggccguuaggccGaa Iaccgug B   CACGGUC U ACGAGAC       14976   HCV.5nc-41 CHz-7 allyl stab1   c s c s c s g s gga cUGAuGaggccguuaggccGaa Iucucgu B   ACGAGAC C UCCCGGG       14977   HCV.5nc-42 CHz-7 allyl stab1   c s c s c s c s ggg cUGAuGaggccguuaggccGaa Igucucg B   CGAGACC U CCCGGGG       14978   HCV.5nc-44 CHz-7 allyl stab1   u s g s c s c s ccg cUGAuGaggccguuaggccGaa Iaggucu B   AGACCUC C CGGGGCA       14979   HCV.5nc-45 CNz-7 allyl stab1   g s u s g s c s ccc cUGAuGaggccguuaggccGaa Igagguc B   GACCUCC C GGGGCAC       14980   HCV.5nc-51 CHz-7 allyl stab1   u s u s g s c s gag cUGAuGaggccguuaggccGaa Iccccgg B   CCGGGGC A CUCGCAA       14981   HCV.5nc-53 CHz-7 allyl stab1   g s c s u s u s gcg cUGAuGaggccguuaggccGaa Iugcccc B   GGGGCAC U CGCAAGC       14982   HCV.5nc-57 CHz-7 allyl stab1   g s g s g s u s gcu cUGAuGaggccguuaggccGaa Icgagug B   CACUCGC A AGCACCC       14983   HCV.5nc-61 CHz-7 allyl stab1   g s a s u s a s ggg cUGAuGaggccguuaggccGaa Icuugcg B   CGCAAGC A CCCUAUC       14984   HCV.5nc-63 CHz-7 allyl stab1   c s u s g s a s uag cUGAuGaggccguuaggccGaa Iugcuug B   CAAGCAC C CUAUCAG       14985   HCV.5nc-64 CHz-7 allyl stab1   c s c s u s g s aua cUGAuGaggccguuaggccGaa Igugcuu B   AAGCACC C UAUCAGG       14986   HCV.5nc-65 CHz-7 allyl stab1   g s c s c s u s gau cUGAuGaggccguuaggccGaa Iggugcu B   AGCACCC U AUCAGGC       14987   HCV.5nc-73 CHz-7 allyl stab1   g s u s g s g s uac cUGAuGaggccguuaggccGaa Iccugau B   AUCAGGC A GUACCAC       14988   HCV.5nc-78 CHz-7 allyl stab1   g s c s c s u s ugu cUGAuGaggccguuaggccGaa Iuacugc B   GCAGUAC C ACAAGGC       14989   HCV.5nc-79 CHz-7 allyl stab1   g s g s c 2 c s uug cUGAuGaggccguuaggccGaa Iguacug B   CAGUACC A CAAGGCC       14990   HCV.5nc-81 CHz-7 allyl stab1   a s a s g s g s ccu cUGAuGaggccguuaggccGaa Iugguac B   GUACCAC A AGGCCUU       14991   HCV.5nc-87 CHz-7 allyl stab1   u s c s g s c s gaa cUGAuGaggccguuaggccGaa Igccuug B   CAAGGCC U UUCGCGA       14992   HCV.5nc-87b CHz-7 allyl stab1   u s u s g s c s gaa cUGAuGaggccguuaggccGaa Igccuug B   CAAGGCC U UUCGCAA       14993   HCV.5nc-101 CHz-7 allyl stab1   c s g s a s g s uag cUGAuGaggccguuaggccGaa Iuugggu B   ACCCAAC A CUACUCG       14994   HCV.5nc-103 CHz-7 allyl stab1   g s c s c s g s agu cUGAuGaggccguuaggccGaa Iuguugg B   CCAACAC U ACUCGGC       14995   HCV.5nc-106 CHz-7 allyl stab1   c s u s a s g s ccg cUGAuGaggccguuaggccGaa Iuagugu B   ACACUAC U CGGCUAG       14996   HCV.5nc-111 CHz-7 allyl stab1   g s a s c s u s gcu cUGAuGaggccguuaggccGaa Iccgagu B   ACUCGGC U AGCAGUC       14997   HCV.5nc-119 CHz-7 allyl stab1   c s c s c s c s gcg cUGAuGaggccguuaggccGaa Iacugcu B   AGCAGUC U CGCGGGG       14998   HCV.5nc-129 CHz-7 allyl stab1   u s u s g s g s gcg cUGAuGaggccguuaggccGaa Icccccg B   CGGGGGC A CGCCCAA       14999   HCV.5nc-163 CHz-7 allyl stab1   c s u s u s u s cuu cUGAuGaggccguuaggccGaa Iaucaac B   GUUGAUC C AAGAAAG       15000   HCV.5nc-163b CHz-7 allyl stab1   c s u s u s u s cuu cUGAuGaggccguuaggccGaa Iauaaac B   GUUUAUC C AAGAAAG       15001   HCV.5nc-164 CHz-7 allyl stab1   c s c s u s u s ucu cUGAuGaggccguuaggccGaa Igaucaa B   UUGAUCC A AGAAAGG       15002   HCV.5nc-164b CHz-7 allyl stab1   c s c s u s u s ucu cUGAuGaggccguuaggccGaa Igauaaa B   UUUAUCC A AGAAAGG       15003   HCV.5nc-193 CHz-7 allyl stab1   g s u s a s c s acc cUGAuGaggccguuaggccGaa Iaauugc B   GCAAUUC C GGUGUAC       15004   HCV.5nc-201 CHz-7 allyl stab1   a s c s c s g s gug cUGAuGaggccguuaggccGaa Iuacacc B   GGUGUAC U CACCGGU       15005   HCV.5nc-203 CHz-7 allyl stab1   g s a s a s c s cgg cUGAuGaggccguuaggccGaa Iaguaca B   UGUACUC A CCGGUUC       15006   HCV.5nc-205 CHz-7 allyl stab1   c s g s g s a s acc cUGAuGaggccguuaggccGaa Iugagua B   UACUCAC C GGUUCCG       15007   HCV.5nc-211 CHz-7 allyl stab1   g s g s u s c s ugc cUGAuGaggccguuaggccGaa Iaaccgg B   CCGGUUC C GCAGACC       15008   HCV.5nc-214 CHz-7 allyl stab1   a s g s u s g s guc cUGAuGaggccguuaggccGaa Icggaac B   GUUCCGC A GACCACU       15009   HCV.5nc-2l8 CHz-7 allyl stab1   c s c s a s u s agu cUGAuGaggccguuaggccGaa Iucugcg B   CGCAGAC C ACUAUGG       15010   HCV.5nc-219 CHz-7 allyl stab1   g s c s c s a s uag cUGAuGaggccguuaggccGaa Igucugc B   GCAGACC A CUAUGGC       15011   HCV.5nc-221 CHz-7 allyl stab1   g s a s g s c s cau cUGAuGaggccguuaggccGaa Iuggucu B   AGACCAC U AUGGCUC       15012   HCV.5nc-227 CHz-7 allyl stab1   c s c s g s g s gag cUGAuGaggccguuaggccGaa Iccauag B   CUAUGGC U CUCCCGG       15013   HCV.5nc-229 CHz-7 allyl stab1   u s c s c s c s ggg cUGAuGaggccguuaggccGaa Iagccau B   AUGGCUC U CCCGGGA       15014   HCV.5nc-231 CHz-7 allyl stab1   c s c s u s c s ccg cUGAuGaggccguuaggccGaa Iagagcc B   GGCUCUC C CGGGAGG       15015   HCV.5nc-232 CHz-7 allyl stab1   c s c s c s u s ccc cUGAuGaggccguuaggccGaa Igagagc B   GCUCUCC C GGGAGGG       15016   HCV.5nc-266 CHz-7 allyl stab1   g s u s u s a s gua cUGAuGaggccguuaggccGaa Iaguguc B   GACACUC A UACUAAC       15017   HCV.5nc-270 CHz-7 allyl stab1   u s g s g s c s guu cUGAuGaggccguuaggccGaa Iuaugag B   CUCAUAC U AACGCCA       15018   HCV.5-31 CHz-7 allyl stab1   u s c s a s c s agg cUGAuGaggccguuaggccGaa Iagugau B   AUCACUC C CCUGUGA       15019   HCV.5-32 CHz-7 allyl stab1   c s u s c s a s cag cUGAuGaggccguuaggccGaa Igaguga B   UCACUCC C CUGUGAG       15020   HCV.5-33 CHz-7 allyl stab1   c s c s u s c s aca cUGAuGaggccguuaggccGaa Iggagug B   CACUCCC C UGUGAGG       15021   HCV.5-34 CHz-7 allyl stab1   u s c s c s u s cac cUGAuGaggccguuaggccGaa Igggagu B   ACUCCCC U GUGAGGA       15022   HCV.5-44 CHz-7 allyl stab1   a s g s a s c s agu cUGAuGaggccguuaggccGaa Iuuccuc B   GAGGAAC U ACUGUCU       15023   HCV.5-47 CHz-7 allyl stab1   u s g s a s a s gac cUGAuGaggccguuaggccGaa Iuaguuc B   GAACUAC U GUCUUCA       15024   HCV.5-51 CHz-7 allyl stab1   u s g s c s g s uga cUGAuGaggccguuaggccGaa Iacagua B   UACUGUC U UCACGCA       15025   HCV.5-54 CHz-7 allyl stab1   u s u s c s u s gcg cUGAuGaggccguuaggccGaa Iaagaca B   UGUCUUC A CGCAGAA       15026   HCV.5-58 CHz-7 allyl stab1   c s g s c s u s uuc cUGAuGaggccguuaggccGaa Icgugaa B   UUCACGC A GAAAGCG       15027   HCV.5-68 CHz-7 allyl stab1   c s a s u s g s gcu cUGAuGaggccguuaggccGaa Iacgcuu B   AAGCGUC U AGCCAUG       15028   HCV.5-72 CHz-7 allyl stab1   a s c s g s c s cau cUGAuGaggccguuaggccGaa Icuagac B   GUCUAGC C AUGGCGU       15029   HCV.5-73 CHz-7 allyl stab1   a s a s c s g s cca cUGAuGaggccguuaggccGaa Igcuaga B   UCUAGCC A UGGCGUU       15030   HCV.5-97 CHz-7 allyl stab1   u s g s g s a s ggc cUGAuGaggccguuaggccGaa Icacgac B   GUCGUGC A GCCUCCA       15031   HCV.5-100 CHz-7 allyl stab1   u s c s c s u s gga cUGAuGaggccguuaggccGaa Icugcac B   GUGCAGC C UCCAGGA       15032   HCV.5-101 CHz-7 allyl stab1   g s u s c s c s ugg cUGAuGaggccguuaggccGaa Igcugca B   UGCAGCC U CCAGGAC       15033   HCV.5-103 CHz-7 allyl stab1   g s g s g s u s ccu cUGAuGaggccguuaggccGaa Iaggcug B   CAGCCUC C AGGACCC       15034   HCV.5-104 CHz-7 allyl stab1   g s g s g s g s ucc cUGAuGaggccguuaggccGaa Igaggcu B   AGCCUCC A GGACCCC       15035   HCV.5-109 CHz-7 allyl stab1   g s a s g s g s ggg cUGAuGaggccguuaggccGaa Iuccugg B   CCAGGAC C CCCCCUC       15036   HCV.5-llO CHz-7 allyl stab1   g s g s a s g s ggg cUGAuGaggccguuaggccGaa Iguccug B   CAGGACC C CCCCUCC       15037   HCV.5-ll1 CHz-7 allyl stab1   g s g s g s a s ggg cUGAuGaggccguuaggccGaa Igguccu B   AGGACCC C CCCUCCC       15038   HCV.5-112 CHz-7 allyl stab1   c s g s g s g s agg cUGAuGaggccguuaggccGaa Igggucc B   GGACCCC C CCUCCCG       15039   HCV.5-113 CHz-7 allyl stab1   c s c s g s g s gag cUGAuGaggccguuaggccGaa Igggguc B   GACCCCC C CUCCCGG       15040   HCV.5-114 CHz-7 allyl stab1   c s c s c s g s gga cUGAuGaggccguuaggccGaa Igggggu B   ACCCCCC C UCCCGGG       15041   HCV.5-115 CHz-7 allyl stab1   u s c s c s c s ggg cUGAuGaggccguuaggccGaa Igggggg B   CCCCCCC U CCCGGGA       15042   HCV.5-117 CHz-7 allyl stab1   u s c s u s c s ccg cUGAuGaggccguuaggccGaa Iaggggg B   CCCCCUC C CGGGAGA       15043   HCV.5-118 CHz-7 allyl stab1   c s u s c s u s ccc cUGAuGaggccguuaggccGaa Igagggg B   CCCCUCC C GGGAGAG       15044   HCV.5-127 CHz-7 allyl stab1   c s c s a s c s uau cUGAuGaggccguuaggccGaa Icucucc B   GGAGAGC C AUAGUGG       15045   HCV.5-128 CHz-7 allyl stab1   a s c s c s a s cua cUGAuGaggccguuaggccGaa Igcucuc B   GAGAGCC A UAGUGGU       15046   HCV.5-137 CHz-7 allyl stab1   g s u s u s c s cgc cUGAuGaggccguuaggccGaa Iaccacu B   AGUGGUC U GCGGAAC       15047   HCV.5-145 CHz-7 allyl stab1   a s c s u s c s acc cUGAuGaggccguuaggccGaa Iuuccgc B   GCGGAAC C GGUGAGU       15048   HCV.5-155 CHz-7 allyl stab1   a s u s u s c s cgg cUGAuGaggccguuaggccGaa Iuacuca B   UGAGUAC A CCGGAAU       15049   HCV.5-157 CHz-7 allyl stab1   c s a s a s u s ucc cUGAuGaggccguuaggccGaa Iuguacu B   AGUACAC C GGAAUUG       15050   HCV.5-181 CHz-7 allyl stab1   c s a s a s g s aaa cUGAuGaggccguuaggccGaa Iacccgg B   CCGGGUC C UUUCUUG       15051   HCV.5-182 CHz-7 allyl stab1   c s c s a s a s gaa cUGAuGaggccguuaggccGaa Igacccg B   CGGGUCC U UUCUUGG       15052   HCV.5-186 CHz-7 allyl stab1   u s g s a s u s cca cUGAuGaggccguuaggccGaa Iaaagga B   UCCUUUC U UGGAUCA