Compositions and methods for inhibiting gene expression of hepatitis B virus

Abstract

The present invention relates to a composition and method for inhibiting the gene expression of hepatitis B virus. The present invention relates to a double-stranded ribonucleic acid (dsRNA) for inhibiting the expression of hepatitis B virus genes. The present invention also relates to a pharmaceutical composition containing the dsRNA or nucleic acid molecule or vector encoding the same and a pharmaceutically acceptable carrier; a method for treating a disease caused by hepatitis B virus infection using the pharmaceutical composition; and a method for inhibiting the expression of hepatitis B virus genes in cells.

Description

[0001] This application is a divisional application. The filing date of the original application is June 28, 2012, the application number is 201610903171.4, and the invention title is "Compositions and Methods for Inhibiting Gene Expression of Hepatitis B Virus". Background Art

[0002] The present invention relates to double-stranded ribonucleic acids (dsRNAs) and their use in mediating RNA interference to inhibit gene expression necessary for the replication and pathogenesis of hepatitis B virus genes, particularly in the inhibition of viral polymerase, surface antigen, e-antigen, and X protein. Additionally, the use of the double-stranded ribonucleic acids in treating or preventing chronic liver diseases / diseases, inflammation, fibrotic disorders, and proliferative diseases such as cancer caused by hepatitis B virus infection is also part of the present invention.

[0003] Hepatitis B virus is a strictly liver tropic double-stranded DNA virus. Although DNA is the genetic material, its replication cycle involves a reverse transcription step to replicate from the pre-genomic RNA into DNA. To complete this essential step, the virus-encoded polymerase has reverse transcriptase activity. Hepatitis B virus is classified as a member of the Hepadnaviruses and belongs to the Hepadnaviridae family. Primary infection of hepatitis B virus in adults causes acute hepatitis with symptoms of organ inflammation, fever, jaundice, and elevated liver transaminases in the blood. Approximately 95% of acute hepatitis cases recover without treatment. Those patients who cannot overcome the viral infection undergo a chronic disease process over the years, with an increased risk of developing cirrhosis or liver cancer. Mother-to-infant transmission of hepatitis B virus from infected mothers to newborn infants also causes chronic hepatitis. The treatment options for chronic hepatitis B virus infection are limited and only lead to complete and sustained remission in some cases. Additional clinical and treatment complications occur in patients co-infected with hepatitis B virus and hepatitis C, hepatitis D, or human immunodeficiency virus.

[0004] Hepatitis B virus is transmitted through blood or blood products, semen, vaginal secretions, or saliva. Drug abuse and sexual intercourse are risky activities and promote the spread of the virus. Contact of damaged mucous epithelial cells with contaminated body fluids may be sufficient to cause infection. The incubation period is 40 - 200 days. The risk of infection is proportional to the number of hepatitis B virus particles transmitted. Infants are often infected mother-to-infant by their hepatitis B virus-carrying mothers, which is a major health problem in endemic areas.

[0005] Approximately two billion people are infected with hepatitis B virus and 400 million are chronic carriers. The highly endemic regions are Africa and Southeast Asia, where 20 - 80% of the infected people are cumulatively present.

[0006] Based on sequence homology, hepatitis B virus is classified into genotypes A - H, and genotypes A - D are the most important ones. Genotype A is common in Northern - Western Europe, the United States, South America, and Central America. Genotypes B and C are mainly in China, Japan, Indonesia, and other East Asian countries. Genotype D is found in Southern Europe, North Africa, and South Africa. The disease progression and response to drug treatment vary among different genotypes.

[0007] The infectious hepatitis B virus particle has a diameter of approximately 42 nm. The outer membrane bilayer contains large, medium, and small surface proteins. The associated hepatocyte receptor for viral surface protein binding and internalization is unknown. Many copies of the core protein form a spherical nucleocapsid structure within the virus particle. Each nucleocapsid carries a partially double - stranded DNA and a viral polymerase as the genetic material.

[0008] After being taken up by hepatocytes, the nucleocapsid is transported to the nucleus and releases the DNA. At this time, DNA strand synthesis is completed and the nicks are repaired to obtain a circular covalently closed (ccc) 3.2 - kb supercoiled DNA. The cccDNA serves as a template for the transcription of four major viral mRNAs, which are 3.5, 2.4, 2.1, and 0.7 kb in length. All mRNAs are 5'-capped and polyadenylated at the 3'-end. There are overlapping sequences at the 3'-ends of all four mRNAs.

[0009] The 3.5 - kb mRNA serves as a template for the core protein and the production of the polymerase. In addition, the same transcript serves as a pre - genomic replication intermediate and allows the viral polymerase to initiate reverse transcription into DNA. The core protein is required for nucleocapsid formation. Additionally, sequential processing activity converts some core proteins into a secreted e - antigen. The abundance of e - antigen in the blood is associated with hepatitis B virus replication in the liver and serves as an important diagnostic marker for monitoring the disease process.

[0010] The 2.4 - and 2.1 - kb mRNAs carry the open reading frames pre - S1, pre - S2, and S for the expression of the viral large, medium, and small surface antigens. The S - antigen is associated with infectious, intact particles. Additionally, the blood of infected patients also contains non - infectious particles that are derived solely from the S - antigen and contain no genomic DNA or polymerase. The function of these particles is not fully understood. The complete and persistent undetectability of the S - antigen in the blood is considered a reliable indicator of hepatitis B virus clearance and thus successful cure.

[0011] The 0.7 kb mRNA encodes protein X. This gene product is important for the efficient transcription of viral genes and also acts as a trans-activator of host cell gene expression. The latter activity seems to be important for hepatocyte transformation in the development of liver cancer.

[0012] Recombinant hepatitis B virus s-antigen is used for vaccination. Injection of three doses of formulated s-antigen on day 1, week 4, and month 6 usually induces sufficient titers of neutralizing antibodies. Vaccinated patients are protected for 10 years or longer. However, there is no therapeutic alternative for this vaccine.

[0013] Due to the high rate of natural remission, patients with acute hepatitis B virus infection cannot be treated. However, those patients with detectable s-antigen, e-antigen, or viral DNA in the blood for more than 6 months are considered to be chronically infected. Nucleoside analogs, which are reverse transcriptase activity inhibitors, are the first choice of treatment for many patients. Lamivudine, Tenofovir, or Entecavir inhibit the replication of hepatitis B virus, sometimes to undetectable levels. Improvement in liver function and reduction of liver inflammation are the most important benefits. However, only a few patients achieve complete and sustained remission after the end of treatment. In addition, hepatitis B virus develops drug resistance during increasing treatment duration. This is particularly difficult for patients co-infected with hepatitis B virus and human immunodeficiency virus. Both viruses are susceptible to nucleoside analog drugs and may develop resistance together.

[0014] The second treatment option is to administer interferon-α. Here, patients receive high doses of interferon-α over a 6-month period. Depending on the viral genotype, up to 50% of chronic infections are curable. However, Asian genotype B has a very low response rate. Co-infection with hepatitis D virus or human immunodeficiency virus renders interferon-α completely ineffective. Patients with symptoms of severe liver damage and severe fibrosis cannot undergo interferon-α treatment.

[0015] Despite significant progress in the field of hepatitis B virus treatment, there is still a need for agents that can selectively and effectively silence viral gene expression, block replication, and continuously reduce viral load in patients with chronic infection.

[0016] Double-stranded RNA molecules (dsRNA) have been shown to block gene expression in a highly conserved regulatory mechanism called RNA interference (RNAi). The present invention provides double-stranded ribonucleic acid molecules (dsRNA), and compositions and methods for inhibiting hepatitis B virus gene expression, particularly for using such dsRNA to inhibit the expression of hepatitis B virus genes in cells, tissues or mammals. The present invention also provides for the treatment or prevention of pathological conditions and diseases caused by hepatitis B virus infection such as chronic liver diseases / conditions, inflammation, fibrotic conditions and proliferative diseases such as cancer. Summary of the Invention

[0017] The present invention provides double-stranded ribonucleic acid (dsRNA) molecules capable of selectively and effectively reducing the expression of hepatitis B virus genes. The use of hepatitis B virus RNAi provides a method for the therapeutic and / or prophylactic treatment of diseases / conditions associated with chronic liver diseases / conditions, inflammation, fibrotic conditions and proliferative diseases such as cancer, such method comprising administering to a human or animal a dsRNA targeting hepatitis B virus.

[0018] In a preferred embodiment, the described dsRNA molecules are capable of inhibiting the expression of at least 60%, preferably at least 70%, most preferably at least 80% of hepatitis B virus genes.

[0019] In one embodiment, the present invention provides double-stranded ribonucleic acid (dsRNA) molecules for inhibiting the expression of hepatitis B virus genes, particularly genes associated with the replication or pathogenic mechanism of hepatitis B. The dsRNA comprises at least two complementary sequences. The dsRNA comprises a sense strand containing a first sequence and an antisense strand containing a second sequence, see the sequences provided in the sequence listing and the specific dsRNA pairs in Tables 1 and 2 attached hereto. In one embodiment, the sense strand comprises a sequence having at least 90% identity with at least a portion of the hepatitis B virus mRNA. Said sequence is located in the complementary region of the sense strand and the antisense strand, preferably within 2-7 nucleotides of the 5'-end of the antisense strand. In a preferred embodiment, the dsRNA specifically targets hepatitis B virus genes encoding core protein, viral polymerase, surface antigen, e-antigen or X protein. Thus, preferably the dsRNA specifically targets a consensus sequence having a highly conserved nucleic acid sequence among hepatitis B virus genomic sequences of genotypes A, B, C and D. Preferably, the consensus sequence is at least 13 contiguous nucleotides in length, more preferably at least 17 contiguous nucleotides, most preferably at least 19 contiguous nucleotides. Preferred highly conserved nucleic acid sequences are shown in Table 5.

[0020] In one embodiment, the antisense strand comprises a region that is substantially complementary to at least a portion of the mRNA encoding the hepatitis B virus gene, and most preferably has a complementary region length of less than 30 nucleotides. Additionally, it is preferred that the length of the dsRNA molecule (double-stranded length) of the invention described herein is in the range of about 16 - 30 nucleotides, particularly in the range of about 18 - 28 nucleotides. Specifically useful in the context of the present invention are double-stranded lengths of about 19, 20, 21, 22, 23, or 24 nucleotides. More preferably, the double-stranded extension is 19, 21, or 23 nucleotides. After delivery to hepatitis B virus-infected cells, the dsRNA inhibits the expression of the hepatitis B virus gene by at least 60%, preferably at least 70%, and most preferably at least 80% in vitro. Brief Description of the Drawings

[0021] Figure 1 . Table 1. Core sequences of dsRNAs targeting the hepatitis B virus gene. Uppercase letters represent RNA nucleotides.

[0022] Figure 2 . Table 2. Characterization of dsRNAs targeting the hepatitis B virus: Activity test of single dose. Uppercase letters represent RNA nucleotides, lowercase letters "c", "g", "a", and "u" represent 2'O-methyl modified nucleotides, "s" represents phosphorothioate, "dT" represents deoxythymidine, and uppercase letters A, C, G, U followed by "f" represent 2'-fluoro nucleotides. Lowercase letter "p" represents 5'-phosphate. (invdT) represents inverted deoxythymidine (3'-3'-linkage).

[0023] Figure 3 . Table 3. Characterization of dsRNAs targeting the hepatitis B virus: Stability t1 / 2 = half-life of the strand as described in the examples.

[0024] Figure 4 . Table 4. Core sequences of dsRNAs targeting the hepatitis B virus gene and their modified counterparts. Uppercase letters represent RNA nucleotides, lowercase letters "c", "g", "a", and "u" represent 2'O-methyl modified nucleotides, "s" represents phosphorothioate, "dT" represents deoxythymidine, and uppercase letters A, C, G, U followed by "f" represent 2'-fluoro nucleotides. Lowercase letter "p" represents 5'-phosphate. (invdT) represents inverted deoxythymidine (3'-3'-linkage).

[0025] Figure 5 . Table 5. Target site sequences of dsRNAs targeting the hepatitis B virus and their coverage relative to hepatitis B virus genotypes A, B, C, and D. n = number of hepatitis B virus sequences obtained for each genotype.

[0026] Figure 6 . Table 6. NCBI GenBank accession numbers of hepatitis B virus genome sequences.

[0027] Figure 7 . Table 7. Comparison of knockdown efficiency and hepatitis B virus genome coverage of individual dsRNAs and their combinations. The activities of two dsRNAs in combination and the best-performing dsRNAs according to Table 2 were tested at final concentrations of 10 nM and 1 nM, and the respective data were compared.

[0028] Figure 8 . Table 8. Sequences of negative control dsRNAs used in the psiCHECK TM -2 screening assay. DETAILED DESCRIPTION OF THE INVENTION

[0029] The attached Table 1 relates to molecules preferably used as dsRNAs according to the present invention. Modified dsRNA molecules are also provided herein and are specifically disclosed in the attached Table 2, providing illustrative examples of the modified dsRNA molecules of the present invention. As noted herein, Table 2 provides illustrative examples of the modified dsRNAs of the present invention (the corresponding sense and antisense strands are provided in this table). The relationship between the unmodified preferred molecules shown in Table 1 and the modified dsRNAs of Table 2 is shown in Table 4. However, the illustrative modifications of these components of the dsRNAs of the present invention provided herein are presented as examples of modifications.

[0030] Table 3 provides selected biological, clinical, and pharmaceutical-related parameters of specific dsRNA molecules of the present invention.

[0031] Some preferred dsRNA molecules are provided in the attached Table 1, and in addition, preferably, the sense strand is selected from the nucleic acid sequences shown in SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, and 26. The antisense strand is selected from the nucleic acid sequences shown in SEQ ID NO: 157, 158, 160, 161, 162, 163, 164, and 186. Thus, the dsRNA molecules of the invention can, in addition, include sequence pairs selected from SEQ ID NO: 1 / 157, 2 / 158, 3 / 160, 4 / 161, 5 / 162, 6 / 163, 7 / 164, and 26 / 186. In the context of the specific dsRNA molecules provided herein, these sequence pairs of SEQ ID NOs relate to the corresponding sense and antisense strand sequences (5'-3') also shown in the table.

[0032] In one embodiment, the dsRNA molecule comprises an antisense strand having a 3' overhang of 1 to 5 nucleotides in length, preferably 1 to 2 nucleotides in length. Preferably, the overhang of the antisense strand comprises uracil or nucleotides complementary to the mRNA encoding a protein necessary for the replication or pathogenesis of hepatitis B virus, especially core protein, viral polymerase, surface antigen, e-antigen and X protein. In another preferred embodiment, the dsRNA molecule comprises a sense strand having a 3' overhang of 1 to 5 nucleotides in length, preferably 1 to 2 nucleotides in length. Preferably, the overhang of the sense strand comprises uracil or nucleotides identical to the mRNA encoding a protein necessary for the replication or pathogenesis of hepatitis B virus.

[0033] In another preferred embodiment, the dsRNA molecule comprises a sense strand having a 3' overhang of 1 to 5 nucleotides in length, preferably 1 to 2 nucleotides in length, and an antisense strand having a 3' overhang of 1 to 5 nucleotides in length, preferably 1 to 2 nucleotides in length. Preferably, the overhang of the sense strand comprises uracil or nucleotides 90% identical to the pregenomic RNA and / or mRNA encoding a protein necessary for the replication or pathogenesis of hepatitis B virus and the overhang of the antisense strand comprises uracil or nucleotides 90% complementary to the mRNA encoding a protein necessary for the replication or pathogenesis of hepatitis B virus.

[0034] The dsDNA molecule of the present invention may comprise naturally occurring nucleotides or may comprise at least one modified nucleotide, such as 2'-O-methyl modified nucleotides, inverted deoxythymidine, nucleotides comprising a 5'-thiophosphate group and terminal nucleotides linked to a cholesteryl derivative or didodecylamid group. 2'-modified nucleotides may have additional advantages in that when the dsRNA molecule of the invention is used in vivo, such as in a medical situation, certain immune stimulatory factors or cytokines are inhibited. Optionally and non-limitingly, the modified nucleotides may be selected from 2'-deoxy-2'-fluoro modified nucleotides, 2'-deoxy modified nucleotides, locked nucleotides, abasic nucleotides, 2'-amino modified nucleotides, 2'-alkyl modified nucleotides, morpholino nucleotides, phosphoramidates and nucleotides containing unnatural bases. In a preferred embodiment, the dsRNA molecule comprises at least one of the following modified nucleotides: 2'-O-methyl modified nucleotides, nucleotides containing a 5'-thiophosphate group and deoxythymidine. Preferred dsRNA molecules comprise modified nucleotides as shown in Table 2. In another preferred embodiment, one of these deoxythymidine nucleotides at the 3' end of both strands is inverted deoxythymidine.

[0035] In a preferred embodiment, the dsRNA molecule of the invention comprises modified nucleotides as detailed in the sequences shown in Table 2. In a preferred embodiment, the dsRNA molecule of the invention comprises sequence pairs selected from SEQ ID NO: 1 / 157, 2 / 158, 3 / 160, 4 / 161, 5 / 162, 6 / 163, 7 / 164, and 26 / 186, and comprises 1-2 deoxythymidines at the overhangs of the antisense strand and / or the sense strand. In a preferred embodiment, the dsRNA molecule of the invention comprises sequence pairs selected from SEQ ID NO: 1 / 157, 2 / 158, 3 / 160, 4 / 161, 5 / 162, 6 / 163, 7 / 164, and 26 / 186, and comprises the modifications detailed in Table 2. Preferred dsRNA molecules comprise the modified nucleotides shown in Tables 2-4, and the most preferred dsRNA molecules are SEQ ID NO: 321 / 485, 322 / 486, 324 / 488, 325 / 489, 326 / 490, 327 / 491, 328 / 492, and 350 / 514.

[0036] In another embodiment, the dsRNA of the invention comprises modified nucleotides at positions different from those shown in Table 2. In a preferred embodiment, two deoxythymidine nucleotides are found at the 3'-ends of both strands of the dsRNA molecule. Preferably, the deoxythymidine nucleotides form overhangs.

[0037] In one embodiment, the dsRNA molecule of the present invention comprises a sense strand and an antisense strand, wherein both strands have a half-life of at least 9 hours. In a preferred embodiment, the dsRNA molecule of the present invention comprises a sense strand and an antisense strand, wherein both strands preferably have a half-life of at least 48 hours in human serum.

[0038] In another embodiment, a nucleic acid sequence encoding the sense strand and / or the antisense strand included in the dsRNA as defined herein is provided.

[0039] The present invention also provides a cell comprising at least one dsRNA of the present invention. The cell is preferably a mammalian cell, such as a human cell. Additionally, a tissue and / or a non-human organism comprising the dsRNA molecule as defined herein is an embodiment of the present invention, wherein the non-human organism is particularly for research purposes or as a research tool, such as in drug testing.

[0040] Additionally, the present invention relates to a method for inhibiting hepatitis B virus gene expression in a cell, tissue, or organism, particularly a hepatitis B virus gene encoding a core protein, a viral polymerase, a surface antigen, an e-antigen, or an X protein, the method comprising the following steps:

[0041] (a) introducing double-stranded ribonucleic acid (dsRNA) as defined herein into a cell, tissue or organism; and

[0042] (b) maintaining the cell, tissue or organism produced in step (a) for a sufficient time to obtain degradation of the mRNA transcript of the hepatitis B virus gene, thereby inhibiting the expression of the hepatitis B virus gene in the designated cell.

[0043] The present invention also relates to a pharmaceutical composition comprising at least one inventive dsRNA. These pharmaceutical compositions are particularly useful for inhibiting the expression of the hepatitis B virus gene in a cell, tissue or organism.

[0044] Preferably, the at least one inventive double-stranded ribonucleic acid molecule targets the pre-genomic RNA or mRNA region encoding a protein necessary for the replication or pathogenesis of the hepatitis B virus gene. More preferably, the targeting region of the inventive double-stranded ribonucleic acid molecule comprises a consensus sequence that is highly conserved among the hepatitis B virus genomic sequences of genotypes A, B, C and D, and the consensus sequence is at least 13 contiguous nucleotides, preferably at least 17 contiguous nucleotides, most preferably at least 19 contiguous nucleotides in length. The preferred highly conserved nucleic acid sequence is shown in Table 5. The pharmaceutical composition can be used to treat patients infected with hepatitis B virus of any genotype or co-infected with different genotypes of hepatitis B virus.

[0045] In the case where the pharmaceutical composition comprises at least two inventive double-stranded ribonucleic acid molecules, preferably the targets of the double-stranded ribonucleic acid molecules are different from each other. The inventive pharmaceutical composition can be used to treat patients and prevent the hepatitis B virus from developing drug resistance to the pharmaceutical composition. In a preferred embodiment, the inventive pharmaceutical composition comprises a combination of dsRNA molecules detailed in the sequences given in Table 7. In a preferred embodiment, the inventive pharmaceutical composition comprises a combination of dsRNA pairs selected from SEQ ID NO: 322 / 486 and 333 / 497, 322 / 486 and 346 / 510, 322 / 486 and 330 / 494, and 322 / 486 and 324 / 488.

[0046] The above-mentioned pharmaceutical composition may also comprise a pharmaceutically acceptable carrier, diluent and / or excipient.

[0047] In another embodiment, the present invention provides a method for treating, preventing or controlling chronic liver diseases / conditions, inflammation, fibrotic conditions and / or proliferative diseases such as cancer associated with hepatitis B virus, the method comprising administering a therapeutically or prophylactically effective amount of one or more inventive dsRNAs to a subject in need of such treatment, prevention or control. Preferably, the subject is a mammal, most preferably a human patient.

[0048] In one embodiment, the present invention provides a method for treating a subject having a pathological condition mediated by hepatitis B virus infection. As described above, such conditions include conditions associated with chronic liver disease / disorder, inflammation, fibrosis symptoms, and / or proliferative disorders such as cancer. In this embodiment, dsRNA is used as a therapeutic agent for controlling the expression of hepatitis B virus genes. The method includes administering the pharmaceutical composition of the present invention to a patient (such as a human) such that the expression of hepatitis B virus genes is silenced. Due to their high specificity, the dsRNAs of the present invention specifically target the mRNA of hepatitis B virus genes. In a preferred embodiment, the dsRNA specifically reduces the level of hepatitis B virus mRNA and does not directly affect the expression and / or mRNA level of off-target genes in the cell.

[0049] In a preferred embodiment, in vivo the dsRNA reduces the level of hepatitis B virus mRNA by at least 60%, preferably at least 70%, and most preferably at least 80% in the liver. In another embodiment, the dsRNA reduces the level of hepatitis B virus mRNA in vivo for at least 4 days. In another preferred embodiment, the dsRNAs of the present invention are used to prepare pharmaceutical compositions for treating chronic liver disease / disorder, inflammation, fibrosis disease, and proliferative diseases such as cancer. Such diseases that require treatment with the pharmaceutical composition include, but are not limited to, chronic hepatitis (CH), liver cirrhosis (HC), and hepatocellular carcinoma (HCC).

[0050] In another embodiment, the present invention provides a vector for inhibiting hepatitis B virus gene expression in a cell, in particular a hepatitis B virus gene comprising a regulatory sequence operably linked to a nucleotide sequence encoding at least one strand of the dsRNA molecule of the present invention.

[0051] In another embodiment, the present invention provides a cell comprising a vector for inhibiting hepatitis B virus gene expression in a cell. The vector comprises a regulatory sequence operably linked to a nucleotide sequence encoding at least one strand of the dsRNA molecule of the present invention. Additionally, in addition to the regulatory sequence, preferably the vector comprises sequences encoding at least one "sense strand" of the dsRNA of the present invention and at least one "antisense strand" of the dsRNA. It is also contemplated that the required cell comprises two or more vectors which, in addition to the regulatory sequence, comprise sequences encoding at least one strand of the dsRNA molecule of the present invention as defined herein.

[0052] In one embodiment, the method includes administering a composition comprising dsRNA, wherein the dsRNA comprises a nucleotide sequence that is complementary to at least a portion of an RNA transcript of a hepatitis B virus gene in a mammal in need of treatment. As noted above, vectors and cells comprising a nucleic acid molecule encoding at least one strand of the dsRNA as defined herein can also be used as pharmaceutical compositions and can therefore also be used in the methods described herein for treating a subject in need of medical intervention. It is also noted that these embodiments of methods involving pharmaceutical compositions and the corresponding subject (human) to be treated also relate to methods such as gene therapy.

[0053] The hepatitis B virus-specific dsRNA molecules provided herein or nucleic acid molecules encoding single strands of these inventive dsRNA molecules can also be inserted into vectors and used as gene therapy vectors for human patients. The gene therapy vectors can be delivered to a subject by, for example, intravenous injection, topical administration (see U.S. Patent No. 5,328,470), or by stereotactic injection (see, e.g., Chen et al., Proc. Natl. Acad. Sci. USA (1994) 91:3054-3057). A pharmaceutical preparation of a gene therapy vector can comprise the gene therapy vector dissolved in an acceptable diluent, or can comprise a slow release formulation in which the gene delivery vehicle is encapsulated. Alternatively, when the intact gene delivery vector can be produced from recombinant cells, such as retroviral vectors, the pharmaceutical preparation can comprise one or more cells that produce the gene delivery system.

[0054] In another aspect of the invention, hepatitis B virus-specific dsRNA molecules that modulate hepatitis B virus gene expression activity are expressed from a transcription unit inserted into a DNA or RNA vector (see, e.g., Skillern A et al., International PCT Publication No. WO 00 / 22113). These transgenes can be introduced as linear constructs, circular plasmids, or viral vectors, which can be incorporated and inherited as transgenes integrated into the host genome. The transgene can be constructed so that it is inherited in the form of an episomal plasmid (Gassmann et al., Proc. Natl. Acad. Sci. USA (1995) 92:1292).

[0055] The single strands of the dsRNA can be transcribed by promoters on two separate expression vectors and co-transfected into the target cells. Optionally, each single strand of the dsRNA can be transcribed by a promoter located on the same expression plasmid. In a preferred embodiment, the dsRNA is expressed as an inverted repeat linked by a linker polynucleotide sequence such that such dsRNA has a stem-loop structure.

[0056] The recombinant dsRNA expression vector is preferably a DNA plasmid or a viral vector. Construction of dsRNA-expressing viral vectors can be based on, but not limited to, adeno-associated virus (for a review, see Muzyczka et al., Curr. Topics Micro. Immunol. (1992) 158:97-129); adenovirus (see, for example, Berkner et al., BioTechniques (1998) 6:616; Rosenfeld et al., Science (1991) 252:431-434; and Rosenfeld et al., Cell (1992) 68:143-155); or alphavirus and other viruses known in the art. Rabies virus has been used to introduce various genes into many different cell types (including epithelial cells) in vitro and / or in vivo (see, for example, Danos and Mulligan, Proc. Natl. Acad. Sci. USA (1998) 85:6460-6464). Recombinant retroviral vectors capable of transducing and expressing genes introduced into the cell genome can be generated by transfection of a recombinant retroviral genome into a suitable packaging cell line such as PA317 and Psi-CRIP (Comette et al., Human Gene Therapy (1991) 2:5-10; Cone et al., Proc. Natl. Acad. Sci. USA (1984) 81:6349). Recombinant adenoviral vectors can be used to infect a variety of cells and tissues of susceptible hosts (such as rats, hamsters, dogs, and gorillas) (Hsu et al., J. Infectious Disease, (1992) 166:769), and also have the advantage of not requiring infection of mitotically active cells.

[0057] The promoter driving dsRNA expression in the DNA plasmid or viral vector of the present invention can be a eukaryotic RNA polymerase I (such as a ribosomal RNA promoter), RNA polymerase II (such as a CMV early promoter or an actin promoter or a U1 snRNA promoter) or preferably an RNA polymerase III promoter (such as a U6 snRNA or 7SK RNA promoter) or a prokaryotic promoter, such as a T7 promoter, provided that the expression plasmid also encodes the T7 RNA polymerase required for transcription from the T7 promoter. The promoter can also direct transgenic expression in the pancreas (see, for example, the insulin regulatory sequences of the pancreas (Bucchini et al., Proc. Natl. Acad. Sci. USA (1986) 83:2511-2515)).

[0058] In addition, for example, by using inducible regulatory sequences and expression systems such as regulatory sequences sensitive to specific physiological regulators (such as circulating glucose levels or hormones), the expression of transgenes can be precisely regulated (Docherty et al., FASEB J. (1994) 8: 20-24). Such inducible expression systems are suitable for controlling transgene expression in cells or mammals, including regulation by ecdysone, estrogen, progesterone, tetracycline, dimerizing chemical inducers, and isopropyl-β-D-1-thiogalactopyranoside (IPTG). Those skilled in the art can select appropriate regulatory / promoter sequences based on the intended use of the dsRNA transgene.

[0059] Preferably, recombinant vectors capable of expressing dsRNA molecules are delivered as described below and persist in target cells. Additionally, viral vectors can be used to provide transient expression of dsRNA molecules. Such vectors can be administered repeatedly as needed. Once expressed, the dsRNA binds to the target RNA and regulates its function or expression. Delivery of dsRNA expression vectors can be systemic, such as by intravenous or intramuscular administration, by administering to target cells transplanted from a patient and then reintroducing them into the patient, or by any other means that allows introduction into the desired target cells.

[0060] dsRNA-expressing DNA plasmids are typically transfected into target cells as complexes with cationic lipid carriers (such as Oligofectamine) or non-cationic lipid-based carriers (such as Transit-TKO TM ). Mixed lipid transfection for dsRNA-mediated knockdown targeting different regions of a single hepatitis B virus gene or multiple hepatitis B virus genes for one week or longer is also within the scope of the present invention. A variety of known methods can be used to monitor the successful introduction of the vectors of the present invention into host cells. For example, transient transfection can be indicated using a reporter gene, such as a fluorescent label (such as green fluorescent protein (GFP)). Markers that confer resistance to specific environmental factors (such as antibiotics or drugs), such as hygromycin B resistance, can be used to ensure stable transfection of isolated cells.

[0061] The following detailed description discloses how to prepare and use dsRNA and compositions containing dsRNA to inhibit the expression of target hepatitis B virus genes, as well as compositions and methods for treating diseases and disorders caused by the hepatitis B virus.

[0062] Definitions

[0063] For convenience, the meanings of specific terms and phrases used in the specification, examples, and appended claims are provided below. If there is an obvious deviation in the use of a term in other parts of this specification from the definition provided in this part, the definition in this part shall prevail.

[0064] "G", "C", "A", "U", and "T" or "dT" each generally represent nucleotides containing guanine, cytosine, adenine, uracil, and deoxythymidine as bases, respectively. However, the terms "ribonucleotide" or "nucleotide" can also refer to modified nucleotides as further defined below, or alternative substitution moieties. Sequences including such substitution moieties are embodiments of the present invention. As detailed below, the dsRNA molecules described herein can also include "overhangs", such as unpaired protruding nucleotides that do not directly participate in the RNA double helix structure, where the RNA double helix structure is generally formed by a pair of "sense strand" and "antisense strand" as defined herein. Such an overhang extension often includes deoxythymidine nucleotides, and in most embodiments, includes two deoxythymidine nucleotides at the 3'-end. Such overhangs are described and illustrated below.

[0065] As used herein, the term "hepatitis B virus gene" refers to the genes necessary for the replication and pathogenesis of hepatitis B virus, especially the genes encoding core protein, viral polymerase, surface antigen, e-antigen, and X protein, and genes encoding functional fragments thereof. The term "hepatitis B virus gene / sequence" refers not only to the wild-type sequence, but also to the mutations and alterations included in the said gene / sequence. Therefore, the present invention is not limited to the specific dsRNA molecules provided herein. The present invention also relates to antisense strand dsRNA molecules including at least 85% complementarity to the nucleotide extension of the RNA transcript of the corresponding hepatitis B virus gene, wherein the hepatitis B virus gene includes such mutations and alterations.

[0066] As described herein, the term "consensus sequence" refers to at least 13 contiguous nucleotides, preferably at least 17 contiguous nucleotides, and most preferably at least 19 contiguous nucleotides, which are highly conserved among the hepatitis B virus genomic sequences of genotypes A, B, C, and D.

[0067] As described herein, a "target sequence" refers to a contiguous portion of the nucleotide sequence of an mRNA molecule formed during the transcription of a hepatitis B virus gene, wherein the mRNA molecule includes the mRNA that is the product of RNA processing of the major transcript.

[0068] As described herein, the term "strand containing a sequence" refers to an oligonucleotide containing a nucleotide chain, the sequence of which is described by using standard nucleotide nomenclature. However, as described herein, such a "strand containing a sequence" can also include modifications, such as modified nucleotides.

[0069] As described herein, and unless otherwise specified, when the term "complementary" is used to describe the relationship between a first nucleotide sequence and a second nucleotide sequence, it refers to the ability of an oligonucleotide or polynucleotide containing the first nucleotide sequence to hybridize with an oligonucleotide or polynucleotide containing the second nucleotide sequence and form a double-stranded structure under specific conditions. As described herein, "complementary" sequences can also include, or consist entirely of, base pairs formed by non-Watson-Crick base pairing and / or non-natural or modified nucleotides, provided that the above requirements regarding their hybridization ability are met.

[0070] Sequences referred to as "fully complementary" include base pairing of an oligonucleotide or polynucleotide containing the first nucleotide sequence with an oligonucleotide or polynucleotide containing the second nucleotide sequence over the entire length of the first nucleotide sequence and the second nucleotide sequence.

[0071] However, in cases where the first sequence is referred to as "substantially complementary" to the second sequence herein, the two sequences can be fully complementary or they can form one or more, but preferably no more than 13, mismatched base pairs upon hybridization.

[0072] The terms "complementary", "fully complementary", and "substantially complementary" herein can be used in reference to base pairing between the sense and antisense strands of a dsRNA, or between the antisense strand of a dsRNA and a target sequence, as can be understood from their context of use.

[0073] As used herein, the terms "double-stranded RNA", "dsRNA molecule" or "dsRNA" refer to ribonucleic acid molecules having a double-stranded structure comprising two anti-parallel and substantially complementary nucleic acid strands, or a complex of ribonucleic acid molecules. The two strands forming the double-stranded structure may be in different regions of a larger RNA molecule or they may be separate RNA molecules. In the case where the two strands are part of a larger molecule and are thus joined by an uninterrupted nucleotide chain between the 3'-end of one strand and the 5'-end of the corresponding other strand forming the double-stranded structure, the joined RNA strands are referred to as a "hairpin loop". In the case where the two strands are covalently joined in a manner other than by an uninterrupted chain between the 3'-end of one strand and the 5'-end of the corresponding other strand forming the double-stranded structure, the joined structure is referred to as a "linker". The RNA strands may have the same or different numbers of nucleotides. Outside the double-stranded structure, the dsRNA may include one or more nucleotide overhangs. The nucleotides in the "overhang" may include from 0 to 5 nucleotides, where "0" indicates no additional nucleotides forming an "overhang" and "5" indicates 5 additional nucleotides on a single strand of the dsRNA duplex. These optional "overhangs" are located at the 3'-end of the single strand. As detailed below, dsRNA molecules including an "overhang" on only one of the two strands can be used and are even advantageous in the context of the present invention. The "overhang" preferably includes from 0 to 2 nucleotides. Most preferably, two "dT" (deoxythymidine) nucleotides are found at the 3'-ends of the two strands of the dsRNA. Two "U" (uracil) can also be in the overhangs at the 3'-ends of the two strands used as dsRNA. Thus, a "nucleotide overhang" refers to unpaired nucleotides or nucleotides protruding from the double-stranded structure of the dsRNA when the 3'-end of one strand of the dsRNA extends beyond the 5'-end of the other strand, or vice versa. For example, an antisense strand comprising 23 nucleotides and a sense strand comprising 21 nucleotides form a two-nucleotide overhang at the 3'-end of the antisense strand. Preferably, the two-nucleotide overhang is fully complementary to the mRNA of the target gene. "Blunt" or "blunt end" refers to no unpaired nucleotides at the ends of the dsRNA, i.e., no nucleotide overhangs. A "blunt end" dsRNA is a dsRNA that is double-stranded throughout its length, i.e., there are no nucleotide overhangs at either end of the molecule.

[0074] The term "antisense strand" refers to the strand of the dsRNA that includes a region substantially complementary to the target sequence. As used herein, the term "complementary region" refers to the region on the antisense strand that is substantially complementary to a sequence, such as the target sequence. In the case where the complementary region is not fully complementary to the target sequence, mismatches are tolerated up to 2 - 7 nucleotides outside the 5'-end of the antisense strand.

[0075] As used herein, the term "sense strand" refers to the strand of a dsRNA that includes a region that is substantially complementary to a region of the antisense strand. "Substantially complementary" means that preferably at least 85% of the overlapping nucleotides in the sense and antisense strands are complementary.

[0076] When "introducing into a cell" refers to a dsRNA, it means facilitating uptake and absorption into the cell, as understood by those skilled in the art. Uptake or absorption of dsRNA can be achieved by simple diffusion or active cellular processes, or by conductive aids or devices. The term is not limited to cells in vitro; dsRNA can also be "introduced into a cell" where the cell is part of a living organism. In such cases, introducing into the cell would include delivery to the organism. For example, for in vivo delivery, dsRNA can be injected into a tissue site or administered systemically. For example, consider the administration of the dsRNA molecules of the present invention to a subject in need of medical intervention. Such administration can include injection of the dsRNA, vector, or cell of the present invention into the diseased site in the subject, such as into liver tissue / cells or into cancerous tissue / cells, such as liver cancer tissue. Additionally, the injection is preferably near the diseased site under consideration. Introduction of cells in vitro includes methods known in the art, such as electroporation and lipofection.

[0077] As used herein, "chronic liver disease / condition" refers to an abnormal liver function that persists for more than 6 months and can be caused by viral infection. An example of a chronic liver disease / condition is chronic hepatitis (CH).

[0078] As used herein, the term "inflammation" refers to the biological response of body tissues to injury, irritation, or disease, which are caused by harmful stimuli such as pathogens, damaged cells, or irritants. Inflammation is typically characterized by pain and swelling. Inflammation includes an acute response, in which inflammatory processes are activated (such as neutrophils and leukocytes), and a chronic response, which is marked by a slow progression, changes in the cell types present at the site of inflammation, and the formation of connective tissue. An example of a disease caused by inflammation is fibrosis.

[0079] As used herein, the term "fibrotic disease" refers to a functional problem of an organ that can be caused by the growth of fibrous tissue. An example of such a disease is hepatic cirrhosis (HC).

[0080] As used herein, the terms "proliferative" and "proliferation" refer to cells that undergo mitosis. In the present application, the term "proliferative disease" refers to any disease / condition characterized by unwanted or abnormal tissue proliferation. The term "proliferative disease" as used herein also refers to a condition in which the growth disorder and / or abnormal growth of cells can lead to the development of an unwanted disorder or disease, which may or may not be cancer.

[0081] Cancers to be treated include, but are not limited to, liver cancer, wherein the liver cancer can be selected from hepatocellular carcinoma (HCC), hepatoblastoma, combined liver cancer, cancers derived from mesenchymal tissues, liver sarcoma, or cholangiocarcinoma.

[0082] The terms "silence", "inhibit expression", and "knockdown", insofar as they refer to hepatitis B virus genes, mean at least partial inhibition of the expression of hepatitis B virus genes, which is manifested as a reduction in the amount of mRNA transcribed from the hepatitis B virus genes, which can be isolated from a first cell or cell population in which the hepatitis B virus genes are transcribed, and compared to a second cell or cell population (control cells) that is substantially the same as the first cell or cell population but has not been so treated, the expression of the hepatitis B virus genes in the first cell or cell population that has been treated is inhibited. The degree of inhibition is usually expressed as

[0083]

[0084] In addition, the degree of inhibition can be counted as a reduction in a parameter that is functionally linked to the transcription of the hepatitis B virus gene, such as the amount of protein encoded by the hepatitis B virus gene secreted by the cell, or the number of cells exhibiting a specific phenotype.

[0085] As shown in the attached examples and the attached tables provided herein, in in vitro tests, the inventive dsRNA molecules are capable of, for example, inhibiting the expression of hepatitis B virus by at least about 60%, preferably at least 70%, and most preferably at least 80% in vitro. The term "in vitro" used herein includes, but is not limited to, cell culture tests. Those skilled in the art can easily determine such inhibition rates and related effects, especially based on the tests provided herein.

[0086] The term "off-target" used herein refers to all non-target mRNAs of the transcriptome that hybridize with the dsRNA predicted by computer methods based on sequence complementarity. The dsRNA of the present invention preferably specifically inhibits the expression of hepatitis B virus genes, that is, it does not inhibit the expression of any off-target genes.

[0087] The term "half-life" used herein is a measure of the stability of a compound or molecule and can be evaluated by methods known to those skilled in the art, especially based on the tests provided herein.

[0088] The term "non-immunostimulatory" used herein means that there is no immune response induced by any of the dsRNA molecules of the present invention. Methods for determining immune responses are well known to those skilled in the art, for example, by evaluating the release of cytokines, as described in the example section.

[0089] The terms "treatment", "therapy", etc. in the context of the present invention refer to the alleviation or mitigation of conditions associated with hepatitis B virus infection, such as chronic liver diseases / conditions, inflammation, fibrotic diseases or proliferative diseases such as cancer.

[0090] As described herein, a "pharmaceutical composition" comprises a pharmaceutically effective amount of at least one dsRNA and a pharmaceutically acceptable carrier. However, such a "pharmaceutical composition" may also comprise single strands of such dsRNA molecules or the carriers described herein, including regulatory sequences operably linked to a nucleotide sequence encoding at least one strand of the sense or antisense strand comprised by the dsRNA of the present invention. Cells, tissues or isolated organs expressing or comprising the dsRNA as defined herein may also be used as a "pharmaceutical composition". As described herein, a "pharmaceutically effective amount", "therapeutically effective amount" or simply "effective amount" refers to the amount of RNA effective to produce the desired pharmacological, therapeutic or prophylactic result.

[0091] The term "pharmaceutically acceptable carrier" refers to a carrier for the administration of a medicament. Such carriers include, but are not limited to, saline, buffered saline, dextrose, water, glycerol, ethanol and combinations thereof. The term specifically excludes cell culture media. For drugs administered orally, pharmaceutically acceptable carriers include, but are not limited to, pharmaceutically acceptable excipients such as inert diluents, disintegrants, binders, lubricants, sweetening agents, flavoring agents, coloring agents and preservatives known to those skilled in the art.

[0092] Particularly contemplated are pharmaceutically acceptable carriers that permit systemic administration of the dsRNA, the carriers of the present invention or the cells. In cases where enteral administration is also contemplated, topical administration and transdermal or transmucosal (such as insufflation, buccal, vaginal, anal) administration and inhalation of the drug are viable ways of administering the compounds of the present invention to a patient in need of medical intervention. When topical administration is employed, this may include direct injection of the compounds of the present invention into the diseased tissue or at least in the vicinity thereof. However, intravenous, intraarterial, subcutaneous, intramuscular, intraperitoneal, intradermal, intrathecal and other modes of administration of the compounds of the present invention are also known to those skilled in the art, such as the attending physician.

[0093] For intramuscular, subcutaneous, and intravenous applications, the pharmaceutical compositions of the invention are generally provided in the form of a sterile aqueous solution or suspension buffered to a suitable pH and conductivity. In a preferred embodiment, the carrier consists entirely of an aqueous buffer. In this context, "entirely" means without conductive adjuvants or encapsulating substances that could affect or mediate the uptake of dsRNA in cells expressing hepatitis B virus genes. The aqueous suspensions according to the invention may include suspending agents such as cellulose derivatives, sodium alginate, polyvinylpyrrolidone, and tragacanth gum, as well as wetting agents such as lecithin. Suitable preservatives for aqueous suspensions include ethyl p-hydroxybenzoate and n-propyl p-hydroxybenzoate. The pharmaceutical compositions useful according to the invention also include encapsulating formulations that protect the dsRNA from rapid elimination from the body, such as controlled release formulations, including implant and microencapsulation delivery systems. Biodegradable, biocompatible polymers such as ethylene vinyl acetate, polyanhydrides, polyglycolic acid, collagen, polyorthoesters, and polylactic acid can be utilized. The methods for preparing such formulations are obvious to those skilled in the art. Liposome suspensions and bispecific antibodies can also be used as pharmaceutically acceptable carriers. These can be prepared by methods known to those skilled in the art, for example, as described in PCT publication numbers WO91 / 06309 and WO2011 / 003780, which are incorporated herein by reference.

[0094] As used herein, a "transformed cell" is a cell into which at least one vector has been introduced, the vector being capable of expressing a dsRNA molecule or at least one strand of such a dsRNA molecule. Such a vector preferably comprises regulatory sequences operably linked to a nucleotide sequence encoding at least one sense or antisense strand of the dsRNA of the invention.

[0095] It can reasonably be expected that shorter dsRNAs comprising one of the sequences in Tables 1 and 4 and having some nucleotides reduced only at one or both ends may have similar effects as compared to the above-mentioned dsRNAs.

[0096] In a preferred embodiment, the dsRNA molecule of the invention comprises nucleotides 1-19 of the sequences shown in Table 1.

[0097] As noted herein, in most embodiments of the invention, the dsRNA molecules provided herein comprise a duplex length of about 16-30 nucleotides (e.g., excluding "overhangs"). Particularly useful dsRNA duplexes have a length of about 19-25 nucleotides. Most preferably, the duplex structure has a length of 19 nucleotides. In the dsRNA molecules of the invention, the antisense strand is at least partially complementary to the sense strand.

[0098] The dsRNA of the present invention can contain one or more mismatches to the target sequence. In a preferred embodiment, the dsRNA of the present invention contains no more than 13 mismatches. If the antisense strand of the dsRNA contains a mismatch to the target sequence, preferably the region of the mismatch is not within 2-7 nucleotides at the 5'-end of the antisense strand. In another embodiment, preferably the region of the mismatch is not within 2-9 nucleotides at the 5'-end of the antisense strand.

[0099] As described above, at least one end / strand of the dsRNA may have 1-5 single-stranded nucleotides, preferably a 1 or 2 nucleotide overhang. The dsRNA with an overhang of at least one nucleotide has unexpectedly excellent inhibitory properties compared to its blunt-ended counterpart. In addition, the inventors have found that the presence of an overhang of only one nucleotide enhances the interfering activity of the dsRNA without affecting its overall stability. The dsRNA with only one overhang has proven to be particularly stable and effective in vivo and in a variety of cells, cell culture media, blood, and serum. Preferably, the single-stranded overhang is located at the 3'-end of the antisense strand or, optionally, at the 3'-end of the sense strand. The dsRNA may also have blunt ends, preferably located at the 5'-end of the antisense strand. Preferably, the antisense strand of the dsRNA has a 1 nucleotide overhang at the 3'-end and the 5'-end is blunt. In another embodiment, one or more of the nucleotides at the overhang are replaced by phosphorothioate nucleotides.

[0100] The dsRNA of the present invention can also be chemically modified to enhance stability. The nucleic acids of the present invention can be synthesized and / or modified by methods well established in the art. Chemical modifications can include, but are not limited to, 2'-modifications, introduction of unnatural bases, covalent attachment to ligands, and replacement of phosphodiester linkages with phosphorothioate linkages, inverted deoxythymidine. In this embodiment, the integrity of the double-stranded structure is strengthened by at least one, and preferably two, chemical linkages. Chemical linkages can be achieved by any of a variety of known techniques, such as by introducing covalent crosslinks, ionic binding or hydrogen bonding; hydrophobic interactions, van der Waals forces or stacking interactions; by means of metal-ion coordination, or by the use of purine analogs. Preferably, chemical groups that can be used to modify dsRNA include, but are not limited to, methylene blue; bifunctional groups, preferably bis-(2-chloroethyl)amine; N-acetyl-N'-(p-glyoxylbenzoyl)cystamine; 4-thiouracil; and psoralen. In a preferred embodiment, the linker is a hexa-ethylene glycol linker. In this case, dsRNA is generated by solid-phase synthesis and the hexa-ethylene glycol linker is incorporated according to standard methods (such as Williams DJ and Hall KB, Biochem. (1996) 35:14665-14670). In a particular embodiment, the 5'-end of the antisense strand and the 3'-end of the sense strand are chemically linked by a hexa-ethylene glycol linker. In another embodiment, at least one nucleotide of the dsRNA comprises a phosphorothioate or dithiophosphate group. The chemical bond at the end of the dsRNA is preferably formed by a triple helix bond.

[0101] In certain embodiments, chemical bonds can be formed by means of one or more bonding groups, where such bonding groups are preferably poly-(oxyphosphinicooxy)-1,3-propanediol and / or polyethylene glycol chains. In other embodiments, chemical bonds can also be formed by a method of introducing purine analogs to replace purines into the double-stranded structure. In additional embodiments, chemical bonds can be formed by introducing azabenzene units into the double-stranded structure. In additional embodiments, chemical bonds are formed by introducing branched nucleotide analogs to replace nucleotides into the double-stranded structure. In certain embodiments, chemical bonds are induced by ultraviolet light.

[0102] In another embodiment, nucleotides that may modify one or both ends of the two single strands are used to block or inhibit the activity of cellular enzymes, such as certain nucleases. Techniques for inhibiting the activity of cellular enzymes are known in the art and include, but are not limited to, 2'-amino modification, 2'-aminosugar modification, 2'-F sugar modification, 2'-F modification, 2'-alkylsugar modification, uncharged backbone modification, morpholino modification, 2'-O-methyl modification, and phosphoramidate (see, e.g., Wagner, Nat. Med. (1995) 1:1116-8). Thus, at least one 2'-hydroxyl group of the nucleotides on the dsRNA is replaced by a chemical group, preferably by 2'-amino or 2'-methyl. Moreover, at least one nucleotide is modified to form a locked nucleotide. Such locked nucleotides contain a methylene bridge that connects the 2'-oxygen of the ribose to the 4'-carbon of the ribose. Introduction of locked nucleotides into oligonucleotides improves the affinity for complementary sequences and raises the melting (unwinding) temperature by several degrees.

[0103] The modifications of the dsRNAs provided herein may positively affect their stability in vivo and in vitro and may also improve their delivery to the (disease) target site. In addition, such structural and chemical modifications may positively affect the physiological response to the dsRNAs after administration, such as preferably inhibited cytokine release. Such chemical and structural modifications are known in the art and are described, inter alia, in Nawrot Current Topics in Med Chem, (2006) 6:913-925.

[0104] Conjugating ligands to dsRNA can enhance its cellular uptake and targeting to specific tissues. In some cases, hydrophobic ligands are conjugated to dsRNA to facilitate its direct passage through the cell membrane. Optionally, the ligand conjugated to the dsRNA is a substrate for receptor-mediated endocytosis. These methods have been used to assist the cellular penetration of antisense oligonucleotides. For example, cholesterol has been conjugated to a variety of antisense oligonucleotides to produce compounds that are more active compared to their unconjugated analogs (see Manoharan M, Antisense & Nucleic Acid Drug Development (2002) 12:103). Other lipophilic compounds conjugated to oligonucleotides include 1-pyrenebutanoic acid, 1,3-di-O-(hexadecyl)glycerol, and menthol. An example of a ligand for receptor-mediated endocytosis is folic acid. Folic acid enters cells through folate receptor-mediated endocytosis. dsRNA compounds with folic acid can be efficiently transported into cells through folate receptor-mediated endocytosis. Binding folic acid to the 3'-end of the oligonucleotide increases the cellular uptake of the nucleotide (Li S, Deshmukh HM, and Huang L, Pharm. Res. (1998) 15:1540). Other ligands conjugated to oligonucleotides include polyethylene glycol, sugar clusters, crosslinkers, porphyrin conjugates, and delivery peptides.

[0105] In some cases, conjugating cationic ligands to oligonucleotides often improves resistance to nucleases. Representative examples of cationic ligands are propylamine and dimethylpropylamine. Interestingly, when cationic ligands are dispersed throughout the oligonucleotide, it has been reported that antisense oligonucleotides retain their high binding affinity for mRNA. See Manoharan M, Antisense & Nucleic Acid Drug Development (2002) 12:103 and cited herein.

[0106] The dsRNA of the conjugate ligand of the present invention can be synthesized by using dsRNA having pendant reactive functionality, such as derived from attaching a linking molecule to the dsRNA. The reactive oligonucleotide can react directly with commercially available ligands, ligands synthesized with any of a variety of protecting groups, or ligands having a linking moiety attached thereto. In some preferred embodiments, the method of the present invention facilitates the synthesis of dsRNA of conjugate ligands by using nucleoside monomers that are appropriately linked to the ligand and further bound to a solid support material. Such ligand-nucleoside conjugates optionally bound to the solid support material can be prepared, according to some preferred embodiments of the method of the present invention, by the reaction of a selected serum-binding ligand with a linking moiety located at the 5'-position of the nucleoside or oligonucleotide. In certain cases, dsRNA having an aralkyl ligand attached to the 3'-end of the dsRNA is prepared by first covalently attaching a monomer building block via a long-chain aminoalkyl to a support of controlled pore glass. Then, nucleotides are attached to the monomer building block already bound to the solid support by standard solid-phase synthesis techniques. The monomer building block can be a nucleoside or other organic compound suitable for solid-phase synthesis.

[0107] The dsRNA used in the conjugates of the present invention can be readily and routinely prepared by known techniques of solid-phase synthesis. It is also known to prepare other oligonucleotides, such as phosphorothioates and alkylated derivatives, using similar techniques.

[0108] Teachings regarding the synthesis of specific modified oligonucleotides can be found in the following U.S. patents: U.S. Patent No. 5,218,105 describes oligonucleotides conjugated to polyamines; U.S. Patent No. 5,541,307 describes oligonucleotides with modified backbones; U.S. Patent No. 5,521,302 describes monomers for the preparation of oligonucleotides with chiral phosphorus linkages; U.S. Patent No. 5,539,082 describes peptide nucleic acids; U.S. Patent No. 5,554,746 describes oligonucleotides with a β-lactam backbone; U.S. Patent No. 5,571,902 describes methods and materials for the synthesis of oligonucleotides; U.S. Patent No. 5,578,718 describes nucleosides with alkylthio groups, where such groups can be used as linkers to any other moiety at various positions of the nucleoside; U.S. Patent No. 5,587,361 describes oligonucleotides with phosphorothioate linkages of high chiral purity; U.S. Patent No. 5,506,351 describes processes for the preparation of 2'-O-alkylguanine nucleosides and related compounds, including 2,6-diaminopurine compounds; U.S. Patent No. 5,587,469 describes oligonucleotides with Ν-2 substituted purines; U.S. Patent No. 5,587,470 describes oligonucleotides with 3-deazapurines; U.S. Patent No. 5,608,046 describes conjugated 4'-demethyl nucleoside analogs; U.S. Patent No. 5,610,289 describes backbone-modified oligonucleotide analogs; U.S. Patent No. 6,262,241 describes methods for the synthesis of 2'-fluoro-oligonucleotides.

[0109] In the conjugated ligand dsRNAs and sequence-specific nucleosides with ligand molecules of the present invention, oligonucleotides and oligonucleosides can be assembled on a suitable oligonucleotide synthesizer using the following materials: standard nucleotide or nucleoside precursors, or nucleotide or nucleoside conjugated precursors that already have a linking moiety, ligand-nucleotide or nucleoside-conjugated precursors that already have a ligand molecule, or building blocks with non-nucleoside ligands.

[0110] When using nucleotide conjugated precursors that already have a linking moiety, the synthesis of the sequence-specific linked nucleosides is typically completed, and then the ligand molecule reacts with the linking moiety to form an oligonucleotide with a conjugated ligand. Oligonucleotide conjugates with various molecules (such as steroids, vitamins, lipids, and reporter molecules) have been previously described (see Manoharan et al., PCT Application No. W093 / 07883). In a preferred embodiment, the oligonucleotides or linked nucleosides of the present invention are synthesized on an automated synthesizer using phosphoramidites from ligand-nucleoside conjugates in addition to the standard and non-standard phosphoramidites that are commercially available and routinely used in oligonucleotide synthesis.

[0111] Incorporation of 2'-O-methyl, 2'-O-ethyl, 2'-O-propyl, 2'-O-allyl, 2'-O-aminoalkyl or 2'-deoxy-2'-fluoro groups into the nucleosides of oligonucleotides confers enhanced hybridization properties to the oligonucleotides. Additionally, oligonucleotides containing a phosphorothioate backbone have enhanced nuclease stability. Thus, the functionalized, linked nucleosides of the present invention can be extended to include a phosphorothioate backbone or one or both of 2'-O-methyl, 2'-O-ethyl, 2'-O-propyl, 2'-O-allyl, 2'-O-aminoalkyl or 2'-deoxy-2'-fluoro groups.

[0112] In certain preferred embodiments, the functionalized nucleoside sequences of the present invention having an amino group at the 5'-terminus are prepared using a DNA synthesizer and are subsequently reacted with the active ester derivative of a selected ligand. Active ester derivatives are well known to those skilled in the art. Representative active esters include N-hydroxysuccinimide esters, tetrafluorophenol esters, pentafluorophenol esters, and pentachlorophenol esters. The reaction of the amino group and the active ester produces an oligonucleotide in which the selected ligand is attached to the 5'-position via a linking group. The amino group at the 5'-terminus can be prepared by using a 5'-amino modifier C6 reagent. In a preferred embodiment, the ligand molecule can be conjugated to the 5'-position of the oligonucleotide by using a ligand-nucleoside phosphoramidite, in which the ligand is directly or indirectly linked to the 5'-hydroxy group via a linker. Such ligand-nucleoside phosphoramidites are typically used at the end of an automated synthesis procedure to provide an oligonucleotide having the conjugated ligand at the 5'-terminus.

[0113] In a preferred embodiment of the method of the present invention, the preparation of the oligonucleotide having the conjugated ligand begins with the selection of a suitable precursor molecule on which the ligand molecule is built. Generally, the precursor is a suitably protected derivative of a commonly used nucleoside. For example, the synthetic precursors for the oligonucleotides having the conjugated ligand of the present invention include, but are not limited to, 2'-aminoalkoxy-5'-ODMT-nucleosides, 2'-6-aminoalkylamino-5'-ODMT-nucleosides, 5'-6-aminoalkoxy-2'-deoxy-nucleosides, 5'-6-aminoalkoxy-2-protected-nucleosides, 3'-6-aminoalkoxy-5'-ODMT-nucleosides, and 3'-aminoalkylamino-5'-ODMT-nucleosides in which the nucleobase portion of the molecule is protected. Methods for synthesizing such protected nucleoside precursors bearing an amino group are known to those of ordinary skill in the art.

[0114] In many instances, protecting groups are used during the preparation of the compounds of the present invention. As used herein, the term "protected" indicates that the designated moiety has a protecting group attached thereto. In some preferred embodiments of the present invention, the compounds comprise one or more protecting groups. A wide variety of protecting groups can be applied in the methods of the present invention. Generally, protecting groups remain inert to the chemical functionality under specific reaction conditions and can be attached or removed from such functionality of the molecule with substantially no damage to the remaining portion of the molecule.

[0115] Commonly used protecting groups, particularly hydroxyl protecting groups, are well known in the art (Greene and Wuts, Protective Groups in Organic Synthesis, Chapter 2, Second Edition, John Wiley & Sons, New York, 1991). Treatment with base selectively removes amino protecting groups that are stable to acid treatment and is used to prepare reactive amino groups that are selectively available for substitution. Examples of such groups are Fmoc and various substituted sulfonylethyl carbamates exemplified by the Nsc group.

[0116] Other amino protecting groups include, but are not limited to, carbamate protecting groups such as 2-trimethylsilylethoxycarbonyl (Teoc), 1-methyl-1-(4-biphenylyl)ethoxycarbonyl (Bpoc), tert-butoxycarbonyl (BOC), allyloxycarbonyl (Alloc), 9-fluorenyl-methoxycarbonyl (Fmoc), and benzyloxycarbonyl (Cbz); amide protecting groups such as formyl, acetyl, trihaloacetyl, benzoyl, and nitrobenzoyl; sulfonamide protecting groups such as 2-nitrobenzenesulfonyl; and imine and cyclic imide protecting groups such as phthalimido and dithiasuccinoyl. Equivalents of these amino protecting groups are also included in the compounds and methods of the present invention.

[0117] Many solid supports are commercially available and it is easy for those skilled in the art to select the solid support to be used in the solid-phase synthesis step. In certain embodiments, a universal support is used. Universal supports well known in the art can be used to prepare oligonucleotides having uncommon or modified nucleotides at the 3'-end of the oligonucleotide. Additionally, it has been reported that when an oligonucleotide is attached to a solid support through a syn-1,2-acetoxyphosphate group that is more susceptible to alkaline hydrolysis, the oligonucleotide can be cleaved from the universal support under milder conditions. See Guzaev Ai and Manoharan MJ. Am. Chem. Soc. (2003) 125:2380.

[0118] The nucleosides are linked by phosphorus-containing and phosphorus-free covalent internucleoside linkages. For identification purposes, such conjugated nucleosides can be characterized as nucleosides with ligands or ligand-nucleoside conjugates. Linked nucleosides having an aralkyl ligand conjugated to the nucleoside in their sequence show enhanced dsRNA activity when compared to similar dsRNA-like compounds without conjugation.

[0119] The conjugated aralkyl-ligand oligonucleotides of the present invention also include conjugates of oligonucleotides and linked nucleosides, wherein the ligand binds directly to the nucleoside or nucleotide without the intervening action of a linker group. The ligand can preferably bind at the carbonyl, amino, and oxy positions of the ligand through a linking group. Typical linking groups can be ester, amide, and carbamate groups.

[0120] Specific examples of modified oligonucleotides for the conjugated ligand oligonucleotides of the present invention preferably include oligonucleotides containing a modified backbone or unnatural internucleoside linkages. As defined herein, oligonucleotides having a modified backbone or unnatural internucleoside linkages include oligonucleotides that retain a phosphorus atom in the backbone and oligonucleotides that do not have a phosphorus atom in the backbone. For the purposes of the present invention, modified oligonucleotides that do not have a phosphorus atom in the sugar backbone are also considered oligonucleosides.

[0121] Specific oligonucleotide chemical modifications are described below. It is not necessary to have uniform modification at all positions of a given compound. Instead, more than one modification can be incorporated into a single dsRNA compound or even into a single nucleotide thereof.

[0122] Preferred modified internucleoside linkages or backbones include, for example, phosphorothioates, chiral phosphorothioates, dithiophosphates, phosphotriesters, aminoalkyl phosphotriesters, methyl and other alkyl phosphates (including 3'-alkenyl phosphates and chiral phosphates), phosphites, phosphoramidates (including 3'-phosphoramidates and aminoalkylamino phosphoramidates), phosphorothioamidates, thioalkyl phosphates, thioalkyl phosphotriesters, and boranophosphates with normal 3'-5' linkages, analogs of these compounds with 2'-5' linkages, and analogs with inverted polarity, wherein adjacent pairs of nucleoside units are linked in the order: 3'-5' to 5'-3' or 2'-5' to 5'-2'. Also included are various salts, mixed salts, and acid-free forms.

[0123] U.S. patents related to the preparation of the above phosphorus-atom-linked ones include, but are not limited to: U.S. Patent Nos. 4,469,863, 5,023,243, 5,264,423, 5,321,131, 5,399,676, 5,405,939, 5,453,496, 5,455,233, and 5,466,677, each of which is incorporated herein by reference.

[0124] Preferred modified internucleoside linkages or backbones (i.e., oligonucleosides) that do not contain phosphorus atoms have backbones formed by short-chain alkyl or cycloalkyl sugar linkages, mixed heteroatom and alkyl or cycloalkyl sugar linkages, or one or more short-chain heteroatom or heterocyclic sugar linkages. These include backbones having morpholino linkages (formed in part from the glycosyl moiety of the nucleoside); siloxane backbones; sulfide, sulfoxide, and sulfone backbones; formyl and thioformyl backbones; methylene formyl and thioformyl backbones; alkenyl-containing backbones; sulfamate backbones; methylene imino and methylene hydrazino backbones; sulfonate and sulfonamide backbones; amide backbones; and other backbones having mixed N, O, S, and CH 2 backbones of the components.

[0125] Representative U.S. patents related to the preparation of the above oligonucleosides include, but are not limited to: U.S. Patent Nos. 5,034,506, 5,214,134, 5,216,141, 5,264,562, 5,466,677, 5,470,967, 5,489,677, 5,602,240, and 5,663,312, each of which is incorporated herein by reference.

[0126] In other preferred oligonucleotide mimetics, both the glycosyl moiety of the nucleotide unit and the internucleoside linkage, i.e., the backbone, are replaced by new groups. The nucleobase units are retained for hybridization with a suitable nucleic acid target compound. Such oligonucleotides, i.e., oligonucleotide mimetics, that have been shown to have excellent hybridization properties are called peptide nucleic acids (PNA). In PNA compounds, the sugar backbone of the oligonucleotide is replaced with an amide-containing backbone, particularly an aminoethylglycine backbone. The nucleobase is retained and is directly or indirectly attached to an atom of the amide moiety of the backbone. Guidance for the preparation of PNA compounds is found, for example, in U.S. Patent No. 5,539,082.

[0127] Some preferred embodiments of the present invention use oligonucleotides having phosphorothioate groups and oligonucleosides having heteroatom backbones, particularly the -CH 2 -NH-O-CH 2 -,-CH 2 -N(CH 3 )-O-CH 2 -[referred to as methylene (methylimino) or MMI backbone], -CH 2 -O-N(CH 3 )-CH 2 -,-CH 2 -N(CH 3 )-N(CH 3 )-CH2 -and -O-N(CH 3 )-CH 2 -CH 2 -[wherein the native phosphodiester backbone is represented as -O-P-O-CH 2 -] and the amide backbone of U.S. Patent No. 5,602,240 cited above. Oligonucleotides having the morpholino backbone structure of U.S. Patent No. 5,034,506 cited above are also preferred.

[0128] The oligonucleotides used in the oligonucleotides of the conjugate ligands of the present invention may additionally or alternatively include nucleobase (abbreviated as "base" in the art) modifiers or alternatives. As used herein, "unmodified" or "native" nucleobases include purine bases: adenine (A) and guanine (G), and pyrimidine bases: thymine (T), cytosine (C), and uracil (U). Modified nucleobases include other synthetic and natural nucleobases such as: 5-methylcytosine (5-me-C), 5-hydroxymethylcytosine, xanthine, hypoxanthine, 2-aminoadenine, 6-methyl and other alkyl derivatives of adenine and guanine, 2-propyl and other alkyl derivatives of adenine and guanine, 2-thiouracil, 2-thiothymine and 2-thiocytosine, 5-halouracil and cytosine, 5-propynyluracil and cytosine, 6-azauracil, cytosine and thymine, 5-uracil (pseudouracil), 4-thiouracil, 8-halo, 8-amino, 8-thio, 8-thioalkyl, 8-hydroxy and other 8-substituted adenines and guanines, 5-halo especially 5-bromo, 5-trifluoromethyl and other 5-substituted uracils and cytosines, 7-methylguanine and 7-methyladenine, 8-azaguanine and 8-azadenine, 7-deazaguanine and 7-deazaadenine, and 3-deazaguanine and 3-deazaadenine.

[0129] Other nucleobases include the nucleobases disclosed in U.S. Patent No. 3,687,808 or other nucleobases known or commercially available in the art. Some of the above nucleobases are particularly useful for enhancing the binding affinity of the oligonucleotides of the present invention. These nucleobases include 5-substituted pyrimidines, 6-azapyrimidines, and N-2, N-6, and O-6 substituted purines, including 2-aminopropyladenine, 5-propynyluracil, and 5-propynylcytosine. It has been found that 5-methylcytosine substitution increases the stability of nucleic acid duplexes by 0.6 - 1.2 °C, which is currently the preferred base substitution and is even more particularly preferred when combined with 2'-methoxyethyl glycosyl modification.

[0130] Representative U.S. patents related to the preparation of some of the above-mentioned modified nucleobases, as well as other modified nucleobases, include, but are not limited to: the above-mentioned U.S. Patent No. 3,687,808, and U.S. Patent Nos. 5,134,066, 5,459,255, 5,552,540, 5,594,121 and 5,596,091, each of which is incorporated herein by reference.

[0131] In certain embodiments, the oligonucleotides used in the oligonucleotides of the conjugated ligands of the present invention may additionally or alternatively include one or more substituted sugar moieties. Preferred oligonucleotides include one of the following at the 2' position: OH, F, O-, S- or N-alkyl, O-, S- or N-alkenyl, or O, S- or N-alkynyl, wherein the alkyl, alkenyl and alkynyl groups may be substituted and unsubstituted C 1 -C 10 Alkyl or C 2 -C 10 Alkenyl and alkynyl. Particularly preferred is O[(CH 2 ) n O] m CH 3 、O(CH 2 ) n OCH 3 、O(CH 2 ) n NH 2 、O(CH 2 ) n CH 3 、O(CH 2 ) n ONH 2 and O(CH 2 ) n ON[(CH 2 ) n CH 3 )] 2 , wherein is 1 to about 10. Other preferred oligonucleotides include the following at the 2' position: C 1 -C 10 Lower alkyl, substituted lower alkyl, alkaryl, aralkyl, O-alkaryl or O-aralkyl, SH, SCH 3 , OCN, Cl, Br, CN, CF 3 、OCF 3 、SOCH 3 、SO 2 CH 3 、ONO 2 、NO 2 、N 3 、NH 2, heterocycloalkyl, heterocycloalkaryl, aminoalkylamino, polyalkylamino, substituted silyl, RNA cleavage group, reporter group, intercalator, group for improving the pharmacokinetic properties of oligonucleotides or group for improving the pharmacodynamic properties of oligonucleotides, and other substituents with similar properties. Preferred modifications include 2'-methoxyethoxy (2'-O-CH 2 CH 2 OCH 3 , also known as 2'-O-(2-methoxyethyl) or 2'-MOE), an alkoxyalkoxy group. Another preferred modification includes 2'-dimethylaminooxyethoxy, i.e., O(CH 2 ) 2 ON(CH 3 ) 2 group, also known as 2'-DMAOE, as described in U.S. Patent No. 6,127,533.

[0132] Other preferred modifications include 2'-methoxy (2'-O-CH 3 ), 2'-aminopropoxy (2'-OCH 2 CH 2 CH 2 NH 2 ), and 2'-fluoro (2'-F). Similar modifications can also be made at other positions of the oligonucleotide, particularly at the 3'-terminal nucleotide or at the 3'-position of the sugar moiety of a 2'-5'-linked oligonucleotide.

[0133] As used herein, the term "sugar substituent" or "2'-substituent" includes a group attached to the 2'-position of a ribofuranosyl moiety with or without an oxygen atom. Sugar substituents include, but are not limited to, fluoro, O-alkyl, O-alkylamino, O-alkylalkoxy, protected O-alkylamino, O-alkylaminoalkyl, O-alkylimidazole, and the formula (O-alkyl) mPolyethers, where m is from 1 to about 10. Preferred among these polyethers are linear and cyclic polyethylene glycols (PEGs) and groups containing PEGs, such as crown ethers and the groups disclosed by Delgardo et al. (Critical Reviews in Therapeutic Drug Carrier Systems (1992) 9:249). Other glycosyl modifications are disclosed by Cook (Anti-fibrosis Drug Design, (1991) 6:585-607). Fluoro, O-alkyl, O-alkylamino, O-alkylimidazole, O-alkylaminoalkyl, and alkylamino substitutions are described in U.S. Patent 6,166,197, entitled "Oligomeric Compounds having Pyrimidine Nucleotide(s) with 2' and 5' Substitutions", the entire content of which is incorporated herein by reference.

[0134] Other sugar substituents suitable for the present invention include 2'-SR and 2'-NR 2 groups, where each R is independently hydrogen, a protecting group, or a substituted or unsubstituted alkyl, alkenyl, or alkynyl group. 2'-SR nucleosides are disclosed in U.S. Patent No. 5,670,633, the entire content of which is incorporated herein by reference. The incorporation of 2'-SR monomer synthetic fibers is disclosed in Hamm et al. (J. Org. Chem., (1997) 62:3415-3420). 2'-NR nucleosides are disclosed in Thomson JB, J. Org. Chem., (1996) 61:6273-6281; and Polushin et al., Tetrahedron Lett., (1996) 37:3227-3230. Other representative 2'-substituted groups suitable for the present invention include groups having one of Formula I or Formula II:

[0135]

[0136] where,

[0137] E is C 1 -C 10 alkyl, N(Q3)(Q4), or N═C(Q3)(Q4); each Q3 and Q4 is independently H, C 1 -C 10an alkyl group, a dialkylaminoalkyl group, a nitrogen protecting group, a tethered or untethered conjugate group, a linker of a solid support; or Q3 and Q4 together form a nitrogen protecting group or a ring structure, which optionally includes at least one additional heteroatom selected from N and O;

[0138] q1 is an integer from 1 to 10;

[0139] q2 is an integer from 1 to 10;

[0140] q3 is 0 or 1;

[0141] q4 is 0, 1 or 2;

[0142] Each of Z1, Z2 and Z3 is independently C 4 -C 7 a cycloalkyl group, C 5 -C 14 an aryl group or C 3 -C 15 a heterocycle, wherein the heteroatoms in the heterocyclic group are selected from oxygen, nitrogen and sulfur;

[0143] Z4 is OM1, SM1 or N(M1) 2 ; each M1 is independently H, C 1 -C 8 an alkyl group, C 1 -C 8 a haloalkyl group, C(=NH)N(H)M2, C(=O)N(H)M2 or OC(=O)N(H)M2; M2 is H or C 1 -C 8 an alkyl group; and

[0144] Z5 is C 1 -C 10 an alkyl group, C 1 -C 10 a haloalkyl group, C 2 -C 10 an alkenyl group, C 2 -C 10 an alkynyl group, C 6 -C 14 an aryl group, N(Q3)(Q4), OQ3, a halogen, SQ3 or CN.

[0145] Representative 2'-O-glycosyl substituents of Formula I are disclosed in U.S. Patent No. 6,172,209, entitled "Capped 2'-Oxyethoxy Oligonucleotides", the entire contents of which are incorporated herein by reference. Representative cyclic 2'-O-glycosyl substituents of Formula II are disclosed in U.S. Patent No. 6,271,358, entitled "RNA Targeted 2'-Modified Oligonucleotides that are Conformationally Preorganized", the entire contents of which are incorporated herein by reference.

[0146] Glycosyls having an O-substituent on the ribose ring are also suitable for the present invention. Representative substituents of the ring O include, but are not limited to, S, CH 2 , CHF, and CF 2 .

[0147] Oligonucleotides may also have sugar mimetics, such as cyclobutyl moieties, which replace the furanose. Representative U.S. patents related to the preparation of such modified glycosyls include, but are not limited to: U.S. Patent Nos. 5,359,044, 5,466,786, 5,519,134, 5,591,722, 5,597,909, 5,646,265, and 5,700,920, each of which is incorporated herein by reference.

[0148] Additional modifications can also be made at other positions of the oligonucleotide, particularly at the 3'-position of the sugar of the 3'-terminal nucleotide. For example, other modifications of the oligonucleotide of the conjugate ligand of the present invention include chemically linking one or more other non-ligand moieties or conjugates to the oligonucleotide, and the non-ligand moieties or conjugates enhance the activity, cellular distribution or cellular uptake of the oligonucleotide. Such moieties include but are not limited to lipid moieties, such as cholesterol moieties (Letsinger et al., Proc. Natl. Acad. Sci. USA, (1989) 86:6553), bile acids (Manoharan et al., Bioorg. Med. Chem. Lett., (1994) 4:1053), thioethers, such as hexyl-S-tritylthiol (Manoharan et al., Ann. N.Y. Acad. Sci., (1992) 660:306; Manoharan et al., Bioorg. Med. Chem. Let., (1993) 3:2765), thiocholesterol (Oberhauser et al., Nucl. Acids Res., (1992) 20:533), aliphatic chains, such as dodecandiol or undecyl residues (Saison-Behmoaras et al., EMBO J., (1991) 10:111; Kabanov et al., FEBS Lett., (1990) 259:327; Svinarchuk et al., Biochimie, (1993) 75:49), phospholipids, such as bis-hexadecyl-rac-glycerol or triethylamine 1,2-bis-O-hexadecyl-rac-glycerol-3-H-phosphonate (Manoharan et al., Tetrahedron Lett., (1995) 36:3651; Shea et al., Nucl. Acids Res., (1990) 18:3777), polyamines or polyethylene glycol chains (Manoharan et al., Nucleosides & Nucleotides, (1995) 14:969), or adamantane acetic acid (Manoharan et al., Tetrahedron Lett., (1995) 36:3651), palmitoyl moieties (Mishra et al., Biochim. Biophys. Acta, (1995) 1264:229), or octadecylamine or hexylamine-carbonyl-oxy cholesterol moieties (Crooke et al., J. Pharmacol. Exp. Ther., (1996) 277:923).

[0149] The invention also includes compositions that use oligonucleotides that are substantially chirally pure at specific positions within the oligonucleotide. Examples of substantially chirally pure oligonucleotides include, but are not limited to, oligonucleotides having phosphorothioate groups that have at least 75% Sp or Rp (Cook et al., U.S. Patent No. 5,587,361), and oligonucleotides having substantially chirally pure (Sp or Rp) alkylphosphonate, aminophosphonate, or phosphotriester linkages (Cook, U.S. Patent Nos. 5,212,295 and 5,521,302).

[0150] In certain instances, the oligonucleotide may be modified by non-ligand groups. A number of non-ligand molecules have been conjugated to oligonucleotides to enhance the activity, cellular distribution, or cellular uptake of the oligonucleotide, and the manner of effecting such conjugations can be found in the scientific literature. Such non-ligand moieties include lipid moieties such as cholesterol moieties (Letsinger et al., Proc. Natl. Acad. Sci. USA, (1989) 86:6553), bile acids (Manoharan et al., Bioorg. Med. Chem. Lett., (1994) 4:1053), thioethers such as hexyl-S-tritylthiol (Manoharan et al., Ann. N.Y. Acad. Sci., (1992) 660:306; Manoharan et al., Bioorg. Med. Chem. Let., (1993) 3:2765), thiocholesterol (Oberhauser et al., Nucl. Acids Res., (1992) 20:533), aliphatic chains such as dodecandiol or undecyl residues (Saison-Behmoaras et al., EMBO J., (1991) 10:111; Kabanov et al., FEBS Lett., (1990) 259:327; Svinarchuk et al., Biochimie, (1993) 75:49), phospholipids such as bis-hexadecyl-rac-glycerol or triethylamine 1,2-bis-O-hexadecyl-rac-glycerol-3-H-phosphonate (Manoharan et al., Tetrahedron Lett., (1995) 36:3651; Shea et al., Nucl. Acids Res., (1990) 18:3777), polyamines or polyethylene glycol chains (Manoharan et al., Nucleosides & Nucleotides, (1995) 14:969), or adamantane acetic acid (Manoharan et al., Tetrahedron Lett., (1995) 36:3651), palmitoyl moieties (Mishra et al., Biochim. Biophys. Acta, (1995) 1264:229), or octadecylamine or hexylamine-carbonyl-oxy cholesterol moieties (Crooke et al., J. Pharmacol. Exp. Ther., (1996) 277:923). Typical conjugation protocols involve synthesizing an oligonucleotide with an amino linker at one or more positions in the sequence. The amino group is then reacted with the molecule to be conjugated using a suitable coupling or activating reaction reagent. The conjugation reaction can be carried out with the oligonucleotide bound to a solid support or in the liquid phase after oligonucleotide cleavage.Purification of oligonucleotide conjugates by HPLC generally results in pure conjugates.

[0151] Optionally, the molecule to be conjugated can be converted into a building block, such as phosphoramidite, which is achieved through an alcohol group in the molecule or by attaching a linker with an alcohol group that can be phosphitylated.

[0152] Importantly, each of these methods can be used to synthesize oligonucleotides conjugated with ligands. The oligonucleotide linking an amino group can be directly conjugated with the ligand using a coupling agent or after activating ligands such as NHS or pentfluorophenolate ester. An aminohexanol linker can be attached to a carboxyl group, and then a ligand phosphoramidite can be synthesized through the phosphite of the terminal alcohol functional group. Other linkers, such as cysteamine, can also be used for conjugation with a chloroacetyl linker in the synthesized oligonucleotide.

[0153] Those skilled in the art can readily conceive of methods for introducing the molecules of the present invention into cells, tissues or organisms. Corresponding examples have also been provided in the above detailed description of the present invention. For example, the nucleic acid molecule or vector of the present invention encoding at least one strand of the dsRNA of the invention can be introduced into cells or tissues by methods known in the art, such as transfection.

[0154] Means and methods for introducing dsRNA molecules are also provided. For example, targeted delivery of glycosylated and folate-modified molecules, including the use of polymeric carriers with ligands such as galactose and lactose or folate-conjugated macromolecules, enables the delivered molecules to bind to folate receptors. Targeted delivery via polypeptides and proteins rather than antibodies is known, for example, including the in vivo delivery of siRNA by RGD-modified nanoparticles or multi-component (non-viral) delivery systems including short cyclodextrins or adamantane-PEG. However, targeted delivery using antibodies or antibody fragments is also contemplated, including the (monovalent) Fab fragments of antibodies (or other fragments of such antibodies) or single-chain antibodies. Injection methods for target-directed delivery include hydrodynamic i.v. injection. Cholesterol conjugates of dsRNA can be used for targeted delivery, whereby conjugation with lipophilic groups enhances cellular uptake and improves the pharmacokinetics and tissue distribution of oligonucleotides. Cationic delivery systems are also known, wherein synthetic carriers bearing a net positive charge (cation) facilitate the formation of complexes with polycationic nucleic acids and interaction with negatively charged cell membranes. Such cationic delivery systems also include cationic liposome delivery systems, cationic polymers, and polypeptide delivery systems. Other delivery systems for cellular uptake of dsRNA / siRNA are aptamer-ds / siRNA. Gene therapy methods can also be used to deliver the inventive dsRNA molecules or nucleic acid molecules encoding the same product. Such systems include the use of non-pathogenic viruses, modified viral vectors, and delivery with nanoparticles or liposomes. Other delivery methods for cellular uptake of dsRNA are ex vivo, such as ex vivo treatment of cells, organs, or tissues. Some such techniques are described and summarized in publications such as Akhtar, Journal of Clinical Investigation (2007) 117:3623-3632, Nguyen et al., Current Opinion in Molecular Therapeutics (2008) 10:158-167, Zamboni, Clin Cancer Res (2005) 11:8230-8234, or Ikeda et al., Pharmaceutical Research (2006) 23:1631-1640.

[0155] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs. Although methods and materials similar or equivalent to those described herein can be used in the practice or testing of the present invention, suitable methods and materials are described below. All publications, patent applications, and patents are incorporated herein by reference in their entirety. In case of conflict, the present specification, including definitions, will control. In addition, the materials, methods, and examples are illustrative only and not intended to be limiting.

[0156] The embodiments and items of the present invention provided above are now illustrated by the following non-limiting examples. Example

[0157] Identification of dsRNA for therapeutic use. dsRNA was designed for the identification of dsRNA specifically targeting hepatitis B virus genotypes A, B, C, and D for therapeutic use.

[0158] First, the known hepatitis B virus genomic sequences (accession numbers shown in Table 6) were downloaded from the NCBI GenBank. Genotype information (accession numbers shown in Table 6) was extracted from the NCBI GenBank file or determined by computer-assisted comparison of the reference genome.

[0159] The genomic sequences of hepatitis B virus genotypes A - D were analyzed by computer to identify optimal target regions for RNAi agents, i.e., 17-nucleotide-long sequence stretches that are highly conserved and identical in at least 90% of all sequences.

[0160] Among the identified RNAi agents, 17-mer sequences were selected by using a specialized algorithm that is limited to having at least two mismatches with any sequence in the human RefSeq database (release 41), and we assume that the human RefSeq database represents the comprehensive human transcriptome.

[0161] All 17-mer sequences containing 4 or more consecutive Gs (poly-G sequences) were further excluded from the synthesis.

[0162] 19-nucleotide-long sequences were defined to include the selected 17-mers at positions 2 - 18.

[0163] These 19-mer sequences cross-react with the genomic sequences of hepatitis B virus genotypes A - D and form the basis for the synthesis of RNA interference (RNAi) agents shown in Tables 1 and 2 attached hereto.

[0164] Synthesis of dsRNA. Oligoribonucleotides were synthesized according to the phosphoramidite technique on solid phase. Depending on the scale of the ABI 394 synthesizer (Applied Biosystems) or AKTA oligopilot 100 (GE Healthcare, Freiburg, Germany) used. The synthesis was carried out on a solid support made of controlled pore glass (CPG, loading 75 μmol / g, from Prime Synthesis, Aston, Pennsylvania, USA). All 2'-modified RNA phosphoramidites and auxiliary reagents were purchased from SAFC (Hamburg, Germany). Specifically, the following 2'-O-methyl phosphoramidites were used: (5'-O-dimethoxytrityl-N 6 -(benzoyl)-2'-O-methyl-adenosine-3'-O-(2-cyanoethyl-N,N-diisopropylamino) phosphoramidite, 5'-O-dimethoxytrityl-N 4 -(acetyl)-2'-O-methyl-cytidine-3'-O-(2-cyanoethyl-N,N-diisopropylamino) phosphoramidite, (5'-O-dimethoxy-trityl-N 2 -(isobutyryl)-2'-O-methyl-guanosine-3'-O-(2-cyanoethyl-N,N-diisopropylamino) phosphoramidite and 5'-O-dimethoxy-trityl-2'-O-methyl-uridine-3'-O-(2-cyanoethyl-N,N-diisopropylamino) phosphoramidite. 2'-Deoxy-2'-fluoro-phosphoramidite carried the same protecting groups as 2'-O-methyl RNA amide. All amidites were dissolved in anhydrous acetonitrile (100 mM) and molecular sieves were added To generate 5'-phosphate, 2-[2-(4,4'-dimethoxytrityloxy)ethylsulfonyl]ethyl-(2-cyanoethyl)-(N,N-diisopropyl)-phosphoramidite from Glen Research, Sterling, Virginia, USA was used. To introduce a C-6 amino linker at the 5'-end of the oligomer, 6-(trifluoroacetylamino)-hexyl-(2-cyanoethyl)-(N,N-diisopropyl)-phosphoramidite from ThermoFisher Scientific, Milwaukee, Wisconsin, USA was used. The 5'-modification was introduced without any modification to the synthesized loop. 5-Ethylthiotetrazole (ETT, 500 mM in acetonitrile) was used as the activator solution. The coupling time was 6 minutes. To introduce phosphorothioate linkages, a 50 mM solution of 3-((dimethylamino-methylene)amino)-3H-1,2,4-dithiazole-3-thione in anhydrous acetonitrile / pyridine (1:1 v / v) (DDTT, obtained from AM Chemicals, Oceanside, CA, USA) was used.

[0165] Cleavage and deprotection of the oligomer bound to the support. After termination of the solid-phase synthesis, the cyanoethyl protecting groups were removed by treatment with 20% diethylamine in ACN for 30 minutes without cleaving the oligonucleotide from the support. Subsequently, the dried solid support was transferred to a 15 ml tube and treated with concentrated ammonia (Aldrich) at 40 °C for 18 hours. After centrifugation, the supernatant was transferred to a new tube and the CPG was washed with ammonia. The combined solutions were evaporated and the solid residue was redissolved in buffer A (see below).

[0166] Purification of the oligoribonucleotide. The crude oligomer was purified by anion-exchange HPLC using Source Q15 (GE Healthcare) and an AKTA detector system (GE Healthcare). Buffer A was 10 mM sodium perchlorate, 20 mM Tris, 1 mM EDTA, pH 7.4 (Fluka Chemie, Buchs, Switzerland) and contained 20% acetonitrile, and buffer B was the same as buffer A except for 500 mM sodium perchlorate. A gradient of 22% B - 42% B was used over 32 column volumes (CV). The UV absorption was recorded at 280 nm, the appropriate fractions were collected and precipitated with 3M NaOAc, pH = 5.2 and 70% ethanol. Finally, the pellet was washed with 70% ethanol. Additionally, desalting was performed using a Sephadex HiTrap column (GE Healthcare) according to the manufacturer's recommendations.

[0167] Annealing of oligoribonucleotides generates siRNA. Complementary strands are mixed by combining equimolar RNA solutions. The mixture is lyophilized and redissolved in an appropriate volume of annealing buffer (100 mM NaCl, 20 mM sodium phosphate, pH 6.8) to achieve the desired concentration. The solution is placed in a water bath at 80 °C and cooled to room temperature over 3 hours.

[0168] In vitro screening of dsRNA targeted to hepatitis B virus mRNA. psiCHECK TM -2 vector (Promega) contains two reporter genes to monitor the activity of RNAi: the Renilla luciferase (hRluc) gene and the synthetic formation of the firefly luciferase gene (hluc+). Measurement of firefly luciferase activity determines changes not related to the activity of RNAi of the tested dsRNA. Using the luciferase assay system (Promega) to measure Renilla luciferase and firefly luciferase activities. After being cloned into the cloning region at the 3' end of the stop codon and polyA tail of the synthetic Renilla luciferase, the hepatitis B virus target site of interest is inserted into psiCHECK TM -2 vector. The cell line COS-7 is transfected with the vector and then treated with dsRNA-lipofectamine 2000 lipid complexes targeting the HBV sequence. The effect of RNAi of dsRNA on the cloned HBV target site is determined by measuring the activity of the Renilla luciferase fusion gene.

[0169] Generation of psiCHECK vectors containing the target sequence. To test the activity of hepatitis B virus dsRNA, a dual-luciferase hepatitis B virus reporter gene was constructed. The 84 - 805, 1075 - 1992, 2165 - 2530, and 2718 - 2940 regions of the hepatitis B virus genome sequence accession number EU554538.1 (genome C) were concatenated by computer. Two mutations (128A→T, 598T→C, relative to the position of EU554538.1) were deliberately inserted. One was needed to remove an internal XhoI site. The second mutation resulted in the removal of a single mismatch for the dsRNA. This hepatitis B virus target construct was extended by adding restriction sites at both the 5' and 3' ends. The artificial DNA sequence was chemically synthesized by Geneart, Regensburg, Germany and cloned into the XhoI / NotI sites of the psiCHECK T M-2 dual-luciferase vector.

[0170] Transfection and luciferase quantification. In 96-well plates at 2.25×10 4Inoculate Cos-7 cells (DSMZ, Braunschweig, Germany, catalog number ACC-60) at a density of cells / well. As described by the manufacturer, plasmid transfection was performed with Lipofectamine 2000 (Invitrogen GmbH, Karlsruhe, Germany, catalog number: 11668-019) at a concentration of 50 ng / well with 0.5 μL / well. Four hours after vector transfection, the medium was discarded and fresh medium was added. After this stage, dsRNA at a concentration of 10 nM or 1 nM was added to the cells using Lipofectamine 2000 as described above. To optimize hepatitis B virus genotype coverage and minimize the development of tolerance to dsRNA, two different dsRNAs can be used in combination simultaneously. To demonstrate the feasibility of such a method, two different dsRNA pairs were selected from the most effective dsRNAs with additional bias to optimize genotype coverage.

[0171] Each dsRNA was added to the cells at a concentration of 5 nM or 0.5 nM to obtain a total dsRNA concentration of 10 nM or 1 nM using Lipofectamine 2000 as described above. As described by the manufacturer, the cells were lysed with luciferase reagent after 48 hours. The Renilla luciferase protein level was normalized relative to the firefly luciferase level to account for transfection efficiency. Four individual data points were collected for each dsRNA. At least one dsRNA not related to all target sites was used as a control to determine the relative Renilla luciferase protein level in cells treated with dsRNA (Table 8). To compare the silencing activity under fully paired conditions, dsRNA fully paired with the Renilla open reading frame was synthesized and tested in parallel with hepatitis B virus dsRNA.

[0172] The inhibition data are given in the attached Table 2.

[0173] Stability of dsRNA. The stability of dsRNA targeting human hepatitis B virus was determined in in vitro assays that measured the half-life of each single strand using any one of human, macaque, or mouse serum.

[0174] 3 μL of a 50 μM dsRNA sample was mixed with 30 μL of human serum (Sigma), rhesus monkey serum (Sigma), or mouse serum (Sigma), and the measurements at each time point were repeated three times. The mixtures were incubated at 37 °C for 0 minutes, 30 minutes, 1 hour, 3 hours, 6 hours, 24 hours, or 48 hours. The reaction was terminated by adding 4 μL of proteinase K (20 mg / ml), 25 μL of "Tissue and Cell Lysis Solution" (Epicentre), and 38 μL of Millipore water and maintaining at 65 °C for 30 minutes. The samples were then filtered by spinning at 1400 rpm through a 0.2 μm 96-well filter plate for 8 minutes, washed twice with 55 μL of Millipore water, and spun-filtered again.

[0175] To isolate the single strands and analyze the remaining full-length product (FLP), the samples were subjected to ion-exchange Dionex Summit HPLC under denaturing conditions, using 20 mM Na 3 PO 4 pH = 11 as eluent A and 1 M NaBr in eluent A as eluent B.

[0176] The following gradient was used:

[0177]

[0178] For each injection, the chromatograms were automatically integrated by Dionex Chromeleon 6.60 HPLC software and, if necessary, adjusted manually. All peak areas were corrected to the internal standard (IS) peak and normalized relative to the incubation at t = 0 minutes. The area under the peak for each single strand and the resulting remaining FLP were calculated and repeated three times. The half-life (t1 / 2) of a strand was defined as the average time point (hours) at which half of the FLP was degraded. The results are given in Table 3 attached. <110> Arrow Pharmaceutical Co., Ltd. <120> Compositions and Methods for Inhibiting Gene Expression of Hepatitis B Virus <130> 27394 WO1 <160> 648 <170> PatentIn version 3.5 <210> 1 <211> 19 <212> RNA <213> Artificial Sequence <223> Artificial Sequence Description: Sense strand of dsRNA <221> Modified Base <222> 1 <223> / Modified base = "Nucleotide: lacking 5'-phosphate group" <400> 1 caagguaugu ugcccguuu 19 <210> 2 <211> 19 <212> RNA <213> Artificial sequence <223> Artificial sequence description: Sense strand of dsRNA <221> Modified base <222> 1 <223> / Modified base = "Nucleotide: lacking 5'-phosphate group" <400> 2 cuguaggcau aaauuggua 19 <210> 3 <211> 19 <212> RNA <213> Artificial sequence <223> Artificial sequence description: Sense strand of dsRNA <221> Modified base <222> 1 <223> / Modified base = "Nucleotide: lacking 5'-phosphate group" <400> 3 ucugcggcgu uuuaucaua 19 <210> 4 <211> 19 <212> RNA <213> Artificial sequence <223> Artificial sequence description: Sense strand of dsRNA <221> Modified base <222> 1 <223> / Modified base = "Nucleotide: lacking 5'-phosphate group" <400> 4 accucugccu aaucaucuc 19 <210> 5 <211> 19 <212> RNA <213> Artificial sequence <223> Artificial sequence description: sense strand of dsRNA <221> Modified base <222> 1 <223> / Modified base = "Nucleotide: lacking 5'-phosphate group" <400> 5 uuuacuagug ccauuugua 19 <210> 6 <211> 19 <212> RNA <213> Artificial sequence <223> Artificial sequence description: sense strand of dsRNA <221> Modified base <222> 1 <223> / Modified base = "Nucleotide: lacking 5'-phosphate group" <400> 6 accucugccu aaucaucua 19 <210> 7 <211> 19 <212> RNA <213> Artificial sequence <223> Artificial sequence description: sense strand of dsRNA <221> Modified base <222> 1 <223> / Modified base = "Nucleotide: lacking 5'-phosphate group" <400> 7 cuguaggcau aaauugguc 19 <210> 8 <211> 19 <212> RNA <213> Artificial sequence <223> Artificial sequence description: sense strand of dsRNA <221> Modified base <222> 1 <223> / Modified base = "Nucleotide: lacking 5'-phosphate group" <400> 8 ugucugcggc guuuuauca 19 <210> 9 <211> 19 <212> RNA <213> Artificial sequence <223> Description of artificial sequence: Sense strand of dsRNA <221> Modified base <222> 1 <223> / Modified base = "Nucleotide: lacking 5'-phosphate group" <400> 9 uacuagugcc auuuguuca 19 <210> 10 <211> 19 <212> RNA <213> Artificial sequence <223> Description of artificial sequence: Sense strand of dsRNA <221> Modified base <222> 1 <223> / Modified base = "Nucleotide: lacking 5'-phosphate group" <400> 10 caacuuuuuc accucugca 19 <210> 11 <211> 19 <212> RNA <213> Artificial sequence <223> Description of artificial sequence: Sense strand of dsRNA <221> Modified base <222> 1 <223> / Modified base = "Nucleotide: lacking 5'-phosphate group" <400> 11 ccauuuguuc agugguucg 19 <210> 12 <211> 19 <212> RNA <213> Artificial sequence <223> Description of artificial sequence: Sense strand of dsRNA <221> Modified base <222> 1 <223> / Modified base = "Nucleotide: lacking 5'-phosphate group" <400> 12 ccaaguguuu gcugacgca 19 <210> 13 <211> 19 <212> RNA <213> Artificial sequence <223> Description of artificial sequence: sense strand of dsRNA <221> Modified base <222> 1 <223> / Modified base = "Nucleotide: lacking 5'-phosphate group" <400> 13 ccauuuguuc agugguuca 19 <210> 14 <211> 19 <212> RNA <213> Artificial sequence <223> Description of artificial sequence: sense strand of dsRNA <221> Modified base <222> 1 <223> / Modified base = "Nucleotide: lacking 5'-phosphate group" <400> 14 uuuacuagug ccauuuguu 19 <210> 15 <211> 19 <212> RNA <213> Artificial sequence <223> Description of artificial sequence: sense strand of dsRNA <221> Modified base <222> 1 <223> / Modified base = "Nucleotide: lacking 5'-phosphate group" <400> 15 caccucugcc uaaucauca 19 <210> 16 <211> 19 <212> RNA <213> Artificial sequence <223> Description of artificial sequence: sense strand of dsRNA <221> Modified base <222> 1 <223> / Modified base = "Nucleotide: lacking 5'-phosphate group" <400> 16 cuggcucagu uuacuagug 19 <210> 17 <211> 19 <212> RNA <213> Artificial sequence <223> Artificial sequence description: sense strand of dsRNA <221> Modified base <222> 1 <223> / Modified base = "Nucleotide: lacking 5'-phosphate group" <400> 17 caagguaugu ugcccguua 19 <210> 18 <211> 19 <212> RNA <213> Artificial sequence <223> Artificial sequence description: sense strand of dsRNA <221> Modified base <222> 1 <223> / Modified base = "Nucleotide: lacking 5'-phosphate group" <400> 18 cuggcucagu uuacuagua 19 <210> 19 <211> 19 <212> RNA <213> Artificial sequence <223> Artificial sequence description: sense strand of dsRNA <221> Modified base <222> 1 <223> / Modified base = "Nucleotide: lacking 5'-phosphate group" <400> 19 gaggcuguag gcauaaauu 19 <210> 20 <211> 19 <212> RNA <213> Artificial sequence <223> Artificial sequence description: sense strand of dsRNA <221> Modified base <222> 1 <223> / Modified base = "Nucleotide: lacking 5'-phosphate group" <400> 20 caguuuacua gugccauuu 19 <210> 21 <211> 19 <212> RNA <213> Artificial sequence <223> Artificial sequence description: Sense strand of dsRNA <221> Modified base <222> 1 <223> / Modified base = "Nucleotide: lacking 5'-phosphate group" <400> 21 agguauguug cccguuugu 19 <210> 22 <211> 19 <212> RNA <213> Artificial sequence <223> Artificial sequence description: Sense strand of dsRNA <221> Modified base <222> 1 <223> / Modified base = "Nucleotide: lacking 5'-phosphate group" <400> 22 uauguugccc guuugucca 19 <210> 23 <211> 19 <212> RNA <213> Artificial sequence <223> Artificial sequence description: Sense strand of dsRNA <221> Modified base <222> 1 <223> / Modified base = "Nucleotide: lacking 5'-phosphate group" <400> 23 gaggcuguag gcauaaaua 19 <210> 24 <211> 19 <212> RNA <213> Artificial sequence <223> Artificial sequence description: sense strand of dsRNA <221> Modified base <222> 1 <223> / Modified base = "Nucleotide: lacking 5'-phosphate group" <400> 24 gucugcggcg uuuuaucau 19 <210> 25 <211> 19 <212> RNA <213> Artificial sequence <223> Artificial sequence description: sense strand of dsRNA <221> Modified base <222> 1 <223> / Modified base = "Nucleotide: lacking 5'-phosphate group" <400> 25 caacuuuuuc accucugcc 19 <210> 26 <211> 19 <212> RNA <213> Artificial sequence <223> Artificial sequence description: sense strand of dsRNA <221> Modified base <222> 1 <223> / Modified base = "Nucleotide: lacking 5'-phosphate group" <400> 26 ccgugugcac uucgcuuca 19 <210> 27 <211> 19 <212> RNA <213> Artificial sequence <223> Artificial sequence description: sense strand of dsRNA <221> Modified base <222> 1 <223> / Modified base = "Nucleotide: lacking 5'-phosphate group" <400> 27 ucaagguaug uugcccgua 19 <210> 28 <211> 19 <212> RNA <213> Artificial sequence <223> Description of artificial sequence: sense strand of dsRNA <221> Modified base <222> 1 <223> / Modified base = "Nucleotide: lacking 5'-phosphate group" <400> 28 caguuuacua gugccauua 19 <210> 29 <211> 19 <212> RNA <213> Artificial sequence <223> Description of artificial sequence: sense strand of dsRNA <221> Modified base <222> 1 <223> / Modified base = "Nucleotide: lacking 5'-phosphate group" <400> 29 ugguggacuu cucucaauu 19 <210> 30 <211> 19 <212> RNA <213> Artificial sequence <223> Description of artificial sequence: sense strand of dsRNA <221> Modified base <222> 1 <223> / Modified base = "Nucleotide: lacking 5'-phosphate group" <400> 30 agguauguug cccguuuga 19 <210> 31 <211> 19 <212> RNA <213> Artificial sequence <223> Description of artificial sequence: sense strand of dsRNA <221> Modified base <222> 1 <223> / Modified base = "Nucleotide: lacking 5'-phosphate group" <400> 31 cugcucgugu uacaggcgg 19 <210> 32 <211> 19 <212> RNA <213> Artificial sequence <223> Artificial sequence description: sense strand of dsRNA <221> Modified base <222> 1 <223> / Modified base = "Nucleotide: lacking 5'-phosphate group" <400> 32 uauguugccc guuuguccu 19 <210> 33 <211> 19 <212> RNA <213> Artificial sequence <223> Artificial sequence description: sense strand of dsRNA <221> Modified base <222> 1 <223> / Modified base = "Nucleotide: lacking 5'-phosphate group" <400> 33 ucaagguaug uugcccguu 19 <210> 34 <211> 19 <212> RNA <213> Artificial sequence <223> Artificial sequence description: sense strand of dsRNA <221> Modified base <222> 1 <223> / Modified base = "Nucleotide: lacking 5'-phosphate group" <400> 34 ucuuaucaac acuuccgga 19 <210> 35 <211> 19 <212> RNA <213> Artificial sequence <223> Artificial sequence description: sense strand of dsRNA <221> Modified base <222> 1 <223> / Modified base = "Nucleotide: lacking 5'-phosphate group" <400> 35 caccucugcc uaaucaucu 19 <210> 36 <211> 19 <212> RNA <213> Artificial sequence <223> Artificial sequence description: Sense strand of dsRNA <221> Modified base <222> 1 <223> / Modified base = "Nucleotide: lacking 5'-phosphate group" <400> 36 auaagaggac ucuuggacu 19 <210> 37 <211> 19 <212> RNA <213> Artificial sequence <223> Artificial sequence description: Sense strand of dsRNA <221> Modified base <222> 1 <223> / Modified base = "Nucleotide: lacking 5'-phosphate group" <400> 37 gucugcggcg uuuuaucaa 19 <210> 38 <211> 19 <212> RNA <213> Artificial sequence <223> Artificial sequence description: Sense strand of dsRNA <221> Modified base <222> 1 <223> / Modified base = "Nucleotide: lacking 5'-phosphate group" <400> 38 ggcgcugaau cccgcggac 19 <210> 39 <211> 19 <212> RNA <213> Artificial sequence <223> Artificial sequence description: Sense strand of dsRNA <221> Modified base <222> 1 <223> / Modified base = "Nucleotide: lacking 5'-phosphate group" <400> 39 cgcgucgcag aagaucuca 19 <210> 40 <211> 19 <212> RNA <213> Artificial sequence <223> Artificial sequence description: Sense strand of dsRNA <221> Modified base <222> 1 <223> / Modified base = "Nucleotide: lacking 5'-phosphate group" <400> 40 aaugucaacg accgaccuu 19 <210> 41 <211> 19 <212> RNA <213> Artificial sequence <223> Artificial sequence description: Sense strand of dsRNA <221> Modified base <222> 1 <223> / Modified base = "Nucleotide: lacking 5'-phosphate group" <400> 41 gcucaguuua cuagugcca 19 <210> 42 <211> 19 <212> RNA <213> Artificial sequence <223> Artificial sequence description: Sense strand of dsRNA <221> Modified base <222> 1 <223> / Modified base = "Nucleotide: lacking 5'-phosphate group" <400> 42 ugguggacuu cucucaaua 19 <210> 43 <211> 19 <212> RNA <213> Artificial sequence <223> Description of artificial sequence: sense strand of dsRNA <221> Modified base <222> 1 <223> / Modified base = "Nucleotide: lacking 5'-phosphate group" <400> 43 aucgccgcgu cgcagaaga 19 <210> 44 <211> 19 <212> RNA <213> Artificial sequence <223> Description of artificial sequence: sense strand of dsRNA <221> Modified base <222> 1 <223> / Modified base = "Nucleotide: lacking 5'-phosphate group" <400> 44 gccauuuguu cagugguuc 19 <210> 45 <211> 19 <212> RNA <213> Artificial sequence <223> Description of artificial sequence: sense strand of dsRNA <221> Modified base <222> 1 <223> / Modified base = "Nucleotide: lacking 5'-phosphate group" <400> 45 cgauccauac ugcggaacu 19 <210> 46 <211> 19 <212> RNA <213> Artificial sequence <223> Description of artificial sequence: sense strand of dsRNA <221> Modified base <222> 1 <223> / Modified base = "Nucleotide: lacking 5'-phosphate group" <400> 46 ucaccucugc cuaaucauc 19 <210> 47 <211> 19 <212> RNA <213> Artificial sequence <223> Description of artificial sequence: sense strand of dsRNA <221> Modified base <222> 1 <223> / Modified base = "Nucleotide: lacking 5'-phosphate group" <400> 47 guggacuucu cucaauuuu 19 <210> 48 <211> 19 <212> RNA <213> Artificial sequence <223> Description of artificial sequence: sense strand of dsRNA <221> Modified base <222> 1 <223> / Modified base = "Nucleotide: lacking 5'-phosphate group" <400> 48 gggucaccau auucuuggg 19 <210> 49 <211> 19 <212> RNA <213> Artificial sequence <223> Description of artificial sequence: sense strand of dsRNA <221> Modified base <222> 1 <223> / Modified base = "Nucleotide: lacking 5'-phosphate group" <400> 49 gccgcgucgc agaagaucu 19 <210> 50 <211> 19 <212> RNA <213> Artificial sequence <223> Description of artificial sequence: sense strand of dsRNA <221> Modified base <222> 1 <223> / Modified base = "Nucleotide: lacking 5'-phosphate group" <400> 50 ucaaucgccg cgucgcaga 19 <210> 51 <211> 19 <212> RNA <213> Artificial sequence <223> Artificial sequence description: Sense strand of dsRNA <221> Modified base <222> 1 <223> / Modified base = "Nucleotide: lacking 5'-phosphate group" <400> 51 uggauguguc ugcggcguu 19 <210> 52 <211> 19 <212> RNA <213> Artificial sequence <223> Artificial sequence description: Sense strand of dsRNA <221> Modified base <222> 1 <223> / Modified base = "Nucleotide: lacking 5'-phosphate group" <400> 52 uacuguucaa gccuccaag 19 <210> 53 <211> 19 <212> RNA <213> Artificial sequence <223> Artificial sequence description: Sense strand of dsRNA <221> Modified base <222> 1 <223> / Modified base = "Nucleotide: lacking 5'-phosphate group" <400> 53 guuuacuagu gccauuugu 19 <210> 54 <211> 19 <212> RNA <213> Artificial sequence <223> Artificial sequence description: Sense strand of dsRNA <221> Modified base <222> 1 <223> / Modified base = "Nucleotide: lacking 5'-phosphate group" <400> 54 acuagugcca uuuguucag 19 <210> 55 <211> 19 <212> RNA <213> Artificial sequence <223> Artificial sequence description: Sense strand of dsRNA <221> Modified base <222> 1 <223> / Modified base = "Nucleotide: lacking 5'-phosphate group" <400> 55 ccgcgucgca gaagaucuc 19 <210> 56 <211> 19 <212> RNA <213> Artificial sequence <223> Artificial sequence description: Sense strand of dsRNA <221> Modified base <222> 1 <223> / Modified base = "Nucleotide: lacking 5'-phosphate group" <400> 56 uaucuuauca acacuuccg 19 <210> 57 <211> 19 <212> RNA <213> Artificial sequence <223> Artificial sequence description: Sense strand of dsRNA <221> Modified base <222> 1 <223> / Modified base = "Nucleotide: lacking 5'-phosphate group" <400> 57 ggccaaaauu cgcaguccc 19 <210> 58 <211> 19 <212> RNA <213> Artificial sequence <223> Artificial sequence description: sense strand of dsRNA <221> Modified base <222> 1 <223> / Modified base = "Nucleotide: lacking 5'-phosphate group" <400> 58 uucaccucug ccuaaucau 19 <210> 59 <211> 19 <212> RNA <213> Artificial sequence <223> Artificial sequence description: sense strand of dsRNA <221> Modified base <222> 1 <223> / Modified base = "Nucleotide: lacking 5'-phosphate group" <400> 59 cucaguuuac uagugccau 19 <210> 60 <211> 19 <212> RNA <213> Artificial sequence <223> Artificial sequence description: sense strand of dsRNA <221> Modified base <222> 1 <223> / Modified base = "Nucleotide: lacking 5'-phosphate group" <400> 60 uguugcccgu uuguccucu 19 <210> 61 <211> 19 <212> RNA <213> Artificial sequence <223> Artificial sequence description: sense strand of dsRNA <221> Modified base <222> 1 <223> / Modified base = "Nucleotide: lacking 5'-phosphate group" <400> 61 uagugccauu uguucagug 19 <210> 62 <211> 19 <212> RNA <213> Artificial sequence <223> Description of artificial sequence: sense strand of dsRNA <221> Modified base <222> 1 <223> / Modified base = "Nucleotide: lacking 5'-phosphate group" <400> 62 aggcuguagg cauaaauug 19 <210> 63 <211> 19 <212> RNA <213> Artificial sequence <223> Description of artificial sequence: sense strand of dsRNA <221> Modified base <222> 1 <223> / Modified base = "Nucleotide: lacking 5'-phosphate group" <400> 63 augugucugc ggcguuuua 19 <210> 64 <211> 19 <212> RNA <213> Artificial sequence <223> Description of artificial sequence: sense strand of dsRNA <221> Modified base <222> 1 <223> / Modified base = "Nucleotide: lacking 5'-phosphate group" <400> 64 acuucgcuuc accucugca 19 <210> 65 <211> 19 <212> RNA <213> Artificial sequence <223> Description of artificial sequence: sense strand of dsRNA <221> Modified base <222> 1 <223> / Modified base = "Nucleotide: lacking 5'-phosphate group" <400> 65 cgugugcacu ucgcuucac 19 <210> 66 <211> 19 <212> RNA <213> Artificial sequence <223> Artificial sequence description: Sense strand of dsRNA <221> Modified base <222> 1 <223> / Modified base=“Nucleotide: lacking 5'-phosphate group” <400> 66 gugguggacu ucucucaau 19 <210> 67 <211> 19 <212> RNA <213> Artificial sequence <223> Artificial sequence description: Sense strand of dsRNA <221> Modified base <222> 1 <223> / Modified base=“Nucleotide: lacking 5'-phosphate group” <400> 67 ugugucugcg gcguuuuau 19 <210> 68 <211> 19 <212> RNA <213> Artificial sequence <223> Artificial sequence description: Sense strand of dsRNA <221> Modified base <222> 1 <223> / Modified base=“Nucleotide: lacking 5'-phosphate group” <400> 68 aagguauguu gcccguuug 19 <210> 69 <211> 19 <212> RNA <213> Artificial sequence <223> Artificial sequence description: Sense strand of dsRNA <221> Modified base <222> 1 <223> / Modified base = "Nucleotide: lacking 5'-phosphate group" <400> 69 ucaacgaccg accuugagg 19 <210> 70 <211> 19 <212> RNA <213> Artificial sequence <223> Description of artificial sequence: sense strand of dsRNA <221> Modified base <222> 1 <223> / Modified base = "Nucleotide: lacking 5'-phosphate group" <400> 70 cauaagagga cucuuggac 19 <210> 71 <211> 19 <212> RNA <213> Artificial sequence <223> Description of artificial sequence: sense strand of dsRNA <221> Modified base <222> 1 <223> / Modified base = "Nucleotide: lacking 5'-phosphate group" <400> 71 gucaacgacc gaccuugag 19 <210> 72 <211> 19 <212> RNA <213> Artificial sequence <223> Description of artificial sequence: sense strand of dsRNA <221> Modified base <222> 1 <223> / Modified base = "Nucleotide: lacking 5'-phosphate group" <400> 72 auauucuugg gaacaagag 19 <210> 73 <211> 19 <212> RNA <213> Artificial sequence <223> Description of artificial sequence: sense strand of dsRNA <221> Modified base <222> 1 <223> / Modified base = "Nucleotide: lacking 5'-phosphate group" <400> 73 ugcucguguu acaggcggg 19 <210> 74 <211> 19 <212> RNA <213> Artificial sequence <223> Artificial sequence description: Sense strand of dsRNA <221> Modified base <222> 1 <223> / Modified base = "Nucleotide: lacking 5'-phosphate group" <400> 74 caaucgccgc gucgcagaa 19 <210> 75 <211> 19 <212> RNA <213> Artificial sequence <223> Artificial sequence description: Sense strand of dsRNA <221> Modified base <222> 1 <223> / Modified base = "Nucleotide: lacking 5'-phosphate group" <400> 75 acuguucaag ccuccaagc 19 <210> 76 <211> 19 <212> RNA <213> Artificial sequence <223> Artificial sequence description: Sense strand of dsRNA <221> Modified base <222> 1 <223> / Modified base = "Nucleotide: lacking 5'-phosphate group" <400> 76 cgccgcgucg cagaagauc 19 <210> 77 <211> 19 <212> RNA <213> Artificial sequence <223> Description of artificial sequence: sense strand of dsRNA <221> Modified base <222> 1 <223> / Modified base = "Nucleotide: lacking 5'-phosphate group" <400> 77 cauuuguuca gugguucgu 19 <210> 78 <211> 19 <212> RNA <213> Artificial sequence <223> Description of artificial sequence: sense strand of dsRNA <221> Modified base <222> 1 <223> / Modified base = "Nucleotide: lacking 5'-phosphate group" <400> 78 cgcugaaucc cgcggacga 19 <210> 79 <211> 19 <212> RNA <213> Artificial sequence <223> Description of artificial sequence: sense strand of dsRNA <221> Modified base <222> 1 <223> / Modified base = "Nucleotide: lacking 5'-phosphate group" <400> 79 ugggucacca uauucuugg 19 <210> 80 <211> 19 <212> RNA <213> Artificial sequence <223> Description of artificial sequence: sense strand of dsRNA <221> Modified base <222> 1 <223> / Modified base = "Nucleotide: lacking 5'-phosphate group" <400> 80 uccucugccg auccauacu 19 <210> 81 <211> 19 <212> RNA <213> Artificial sequence <223> Description of artificial sequence: sense strand of dsRNA <221> Modified base <222> 1 <223> / Modified base = "Nucleotide: lacking 5'-phosphate group" <400> 81 augucaacga ccgaccuug 19 <210> 82 <211> 19 <212> RNA <213> Artificial sequence <223> Description of artificial sequence: sense strand of dsRNA <221> Modified base <222> 1 <223> / Modified base = "Nucleotide: lacking 5'-phosphate group" <400> 82 ccucugccua aucaucuca 19 <210> 83 <211> 19 <212> RNA <213> Artificial sequence <223> Description of artificial sequence: sense strand of dsRNA <221> Modified base <222> 1 <223> / Modified base = "Nucleotide: lacking 5'-phosphate group" <400> 83 accgugugca cuucgcuuc 19 <210> 84 <211> 19 <212> RNA <213> Artificial sequence <223> Description of artificial sequence: sense strand of dsRNA <221> Modified base <222> 1 <223> / Modified base = "Nucleotide: lacking 5'-phosphate group" <400> 84 ugccgaucca uacugcgga 19 <210> 85 <211> 19 <212> RNA <213> Artificial sequence <223> Description of artificial sequence: sense strand of dsRNA <221> Modified base <222> 1 <223> / Modified base = "Nucleotide: lacking 5'-phosphate group" <400> 85 cagagucuag acucguggu 19 <210> 86 <211> 19 <212> RNA <213> Artificial sequence <223> Description of artificial sequence: sense strand of dsRNA <221> Modified base <222> 1 <223> / Modified base = "Nucleotide: lacking 5'-phosphate group" <400> 86 cuguucaagc cuccaagcu 19 <210> 87 <211> 19 <212> RNA <213> Artificial sequence <223> Description of artificial sequence: sense strand of dsRNA <221> Modified base <222> 1 <223> / Modified base = "Nucleotide: lacking 5'-phosphate group" <400> 87 ggaggcugua ggcauaaau 19 <210> 88 <211> 19 <212> RNA <213> Artificial sequence <223> Description of artificial sequence: sense strand of dsRNA <221> Modified base <222> 1 <223> / Modified base = "Nucleotide: lacking 5'-phosphate group" <400> 88 aggaggcugu aggcauaaa 19 <210> 89 <211> 19 <212> RNA <213> Artificial sequence <223> Artificial sequence description: Sense strand of dsRNA <221> Modified base <222> 1 <223> / Modified base = "Nucleotide: lacking 5'-phosphate group" <400> 89 gguggacuuc ucucaauuu 19 <210> 90 <211> 19 <212> RNA <213> Artificial sequence <223> Artificial sequence description: Sense strand of dsRNA <221> Modified base <222> 1 <223> / Modified base = "Nucleotide: lacking 5'-phosphate group" <400> 90 gcaacuuuuu caccucugc 19 <210> 91 <211> 19 <212> RNA <213> Artificial sequence <223> Artificial sequence description: Sense strand of dsRNA <221> Modified base <222> 1 <223> / Modified base = "Nucleotide: lacking 5'-phosphate group" <400> 91 cugcucgugu uacaggcga 19 <210> 92 <211> 19 <212> RNA <213> Artificial sequence <223> Artificial Sequence Description: Sense Strand of dsRNA <221> Modified Base <222> 1 <223> / Modified Base = "Nucleotide: Lacking 5'-phosphate group" <400> 92 cuagugccau uuguucagu 19 <210> 93 <211> 19 <212> RNA <213> Artificial Sequence <223> Artificial Sequence Description: Sense Strand of dsRNA <221> Modified Base <222> 1 <223> / Modified Base = "Nucleotide: Lacking 5'-phosphate group" <400> 93 cugccgaucc auacugcgg 19 <210> 94 <211> 19 <212> RNA <213> Artificial Sequence <223> Artificial Sequence Description: Sense Strand of dsRNA <221> Modified Base <222> 1 <223> / Modified Base = "Nucleotide: Lacking 5'-phosphate group" <400> 94 gugugcacuu cgcuucacc 19 <210> 95 <211> 19 <212> RNA <213> Artificial Sequence <223> Artificial Sequence Description: Sense Strand of dsRNA <221> Modified Base <222> 1 <223> / Modified Base = "Nucleotide: Lacking 5'-phosphate group" <400> 95 gcucguguua caggcgggc 19 <210> 96 <211> 19 <212> RNA <213> Artificial sequence <223> Description of artificial sequence: Sense strand of dsRNA <221> Modified base <222> 1 <223> / Modified base = "Nucleotide: lacking 5'-phosphate group" <400> 96 ccuaucuuau caacacuuc 19 <210> 97 <211> 19 <212> RNA <213> Artificial sequence <223> Description of artificial sequence: Sense strand of dsRNA <221> Modified base <222> 1 <223> / Modified base = "Nucleotide: lacking 5'-phosphate group" <400> 97 ucucaaucgc cgcgucgca 19 <210> 98 <211> 19 <212> RNA <213> Artificial sequence <223> Description of artificial sequence: Sense strand of dsRNA <221> Modified base <222> 1 <223> / Modified base = "Nucleotide: lacking 5'-phosphate group" <400> 98 gcccgucugu gccuucuca 19 <210> 99 <211> 19 <212> RNA <213> Artificial sequence <223> Description of artificial sequence: Sense strand of dsRNA <221> Modified base <222> 1 <223> / Modified base = "Nucleotide: lacking 5'-phosphate group" <400> 99 cuaucuuauc aacacuucc 19 <210> 100 <211> 19 <212> RNA <213> Artificial sequence <223> Artificial sequence description: Sense strand of dsRNA <221> Modified base <222> 1 <223> / Modified base = "Nucleotide: lacking 5'-phosphate group" <400> 100 auguugcccg uuuguccuc 19 <210> 101 <211> 19 <212> RNA <213> Artificial sequence <223> Artificial sequence description: Sense strand of dsRNA <221> Modified base <222> 1 <223> / Modified base = "Nucleotide: lacking 5'-phosphate group" <400> 101 guauguugcc cguuugucc 19 <210> 102 <211> 19 <212> RNA <213> Artificial sequence <223> Artificial sequence description: Sense strand of dsRNA <221> Modified base <222> 1 <223> / Modified base = "Nucleotide: lacking 5'-phosphate group" <400> 102 cuucgcuuca ccucugcac 19 <210> 103 <211> 19 <212> RNA <213> Artificial sequence <223> Artificial sequence description: Sense strand of dsRNA <221> Modified base <222> 1 <223> / Modified base = "Nucleotide: lacking 5'-phosphate group" <400> 103 ugugcacuuc gcuucaccu 19 <210> 104 <211> 19 <212> RNA <213> Artificial sequence <223> Artificial sequence description: sense strand of dsRNA <221> Modified base <222> 1 <223> / Modified base = "Nucleotide: lacking 5'-phosphate group" <400> 104 gccaaaauuc gcagucccg 19 <210> 105 <211> 19 <212> RNA <213> Artificial sequence <223> Artificial sequence description: sense strand of dsRNA <221> Modified base <222> 1 <223> / Modified base = "Nucleotide: lacking 5'-phosphate group" <400> 105 ccugcucgug uuacaggcg 19 <210> 106 <211> 19 <212> RNA <213> Artificial sequence <223> Artificial sequence description: sense strand of dsRNA <221> Modified base <222> 1 <223> / Modified base = "Nucleotide: lacking 5'-phosphate group" <400> 106 uggagugugg auucgcacu 19 <210> 107 <211> 19 <212> RNA <213> Artificial sequence <223> Artificial sequence description: sense strand of dsRNA <221> Modified base <222> 1 <223> / Modified base = "Nucleotide: lacking 5'-phosphate group" <400> 107 aacgaccgac cuugaggca 19 <210> 108 <211> 19 <212> RNA <213> Artificial sequence <223> Artificial sequence description: sense strand of dsRNA <221> Modified base <222> 1 <223> / Modified base = "Nucleotide: lacking 5'-phosphate group" <400> 108 acagagucua gacucgugg 19 <210> 109 <211> 19 <212> RNA <213> Artificial sequence <223> Artificial sequence description: sense strand of dsRNA <221> Modified base <222> 1 <223> / Modified base = "Nucleotide: lacking 5'-phosphate group" <400> 109 aaucgccgcg ucgcagaag 19 <210> 110 <211> 19 <212> RNA <213> Artificial sequence <223> Artificial sequence description: sense strand of dsRNA <221> Modified base <222> 1 <223> / Modified base = "Nucleotide: lacking 5'-phosphate group" <400> 110 gguauguugc ccguuuguc 19 <210> 111 <211> 19 <212> RNA <213> Artificial sequence <223> Description of artificial sequence: sense strand of dsRNA <221> Modified base <222> 1 <223> / Modified base = "Nucleotide: lacking 5'-phosphate group" <400> 111 gccgauccau acugcggaa 19 <210> 112 <211> 19 <212> RNA <213> Artificial sequence <223> Description of artificial sequence: sense strand of dsRNA <221> Modified base <222> 1 <223> / Modified base = "Nucleotide: lacking 5'-phosphate group" <400> 112 gcccuaucuu aucaacacu 19 <210> 113 <211> 19 <212> RNA <213> Artificial sequence <223> Description of artificial sequence: sense strand of dsRNA <221> Modified base <222> 1 <223> / Modified base = "Nucleotide: lacking 5'-phosphate group" <400> 113 aguuuacuag ugccauuug 19 <210> 114 <211> 19 <212> RNA <213> Artificial sequence <223> Description of artificial sequence: sense strand of dsRNA <221> Modified base <222> 1 <223> / Modified base = "Nucleotide: lacking 5'-phosphate group" <400> 114 ugucaacgac cgaccuuga 19 <210> 115 <211> 19 <212> RNA <213> Artificial sequence <223> Description of artificial sequence: Sense strand of dsRNA <221> Modified base <222> 1 <223> / Modified base = "Nucleotide: lacking 5'-phosphate group" <400> 115 acuucucuca auuuucuag 19 <210> 116 <211> 19 <212> RNA <213> Artificial sequence <223> Description of artificial sequence: Sense strand of dsRNA <221> Modified base <222> 1 <223> / Modified base = "Nucleotide: lacking 5'-phosphate group" <400> 116 gcgcgggacg uccuuuguc 19 <210> 117 <211> 19 <212> RNA <213> Artificial sequence <223> Description of artificial sequence: Sense strand of dsRNA <221> Modified base <222> 1 <223> / Modified base = "Nucleotide: lacking 5'-phosphate group" <400> 117 ucuagacucg ugguggacu 19 <210> 118 <211> 19 <212> RNA <213> Artificial sequence <223> Description of artificial sequence: Sense strand of dsRNA <221> Modified base <222> 1 <223> / Modified base = "Nucleotide: lacking 5'-phosphate group" <400> 118 gauccauacu gcggaacuc 19 <210> 119 <211> 19 <212> RNA <213> Artificial sequence <223> Artificial sequence description: Sense strand of dsRNA <221> Modified base <222> 1 <223> / Modified base = "Nucleotide: lacking 5'-phosphate group" <400> 119 cucugccgau ccauacugc 19 <210> 120 <211> 19 <212> RNA <213> Artificial sequence <223> Artificial sequence description: Sense strand of dsRNA <221> Modified base <222> 1 <223> / Modified base = "Nucleotide: lacking 5'-phosphate group" <400> 120 ucugccgauc cauacugcg 19 <210> 121 <211> 19 <212> RNA <213> Artificial sequence <223> Artificial sequence description: Sense strand of dsRNA <221> Modified base <222> 1 <223> / Modified base = "Nucleotide: lacking 5'-phosphate group" <400> 121 ccucugccga uccauacug 19 <210> 122 <211> 19 <212> RNA <213> Artificial sequence <223> Artificial sequence description: sense strand of dsRNA <221> Modified base <222> 1 <223> / Modified base = "Nucleotide: lacking 5'-phosphate group" <400> 122 gcaccucucu uuacgcggu 19 <210> 123 <211> 19 <212> RNA <213> Artificial sequence <223> Artificial sequence description: sense strand of dsRNA <221> Modified base <222> 1 <223> / Modified base = "Nucleotide: lacking 5'-phosphate group" <400> 123 aagaacuccc ucgccucgc 19 <210> 124 <211> 19 <212> RNA <213> Artificial sequence <223> Artificial sequence description: sense strand of dsRNA <221> Modified base <222> 1 <223> / Modified base = "Nucleotide: lacking 5'-phosphate group" <400> 124 gaacucccuc gccucgcag 19 <210> 125 <211> 19 <212> RNA <213> Artificial sequence <223> Artificial sequence description: sense strand of dsRNA <221> Modified base <222> 1 <223> / Modified base = "Nucleotide: lacking 5'-phosphate group" <400> 125 ucucucaauu uucuagggc 19 <210> 126 <211> 19 <212> RNA <213> Artificial sequence <223> Description of artificial sequence: sense strand of dsRNA <221> Modified base <222> 1 <223> / Modified base = "Nucleotide: lacking 5'-phosphate group" <400> 126 gggcgcaccu cucuuuacg 19 <210> 127 <211> 19 <212> RNA <213> Artificial sequence <223> Description of artificial sequence: sense strand of dsRNA <221> Modified base <222> 1 <223> / Modified base = "Nucleotide: lacking 5'-phosphate group" <400> 127 ccgauccaua cugcggaac 19 <210> 128 <211> 19 <212> RNA <213> Artificial sequence <223> Description of artificial sequence: sense strand of dsRNA <221> Modified base <222> 1 <223> / Modified base = "Nucleotide: lacking 5'-phosphate group" <400> 128 aacucccucg ccucgcaga 19 <210> 129 <211> 19 <212> RNA <213> Artificial sequence <223> Description of artificial sequence: sense strand of dsRNA <221> Modified base <222> 1 <223> / Modified base = "Nucleotide: lacking 5'-phosphate group" <400> 129 cuccucugcc gauccauac 19 <210> 130 <211> 19 <212> RNA <213> Artificial sequence <223> Description of artificial sequence: sense strand of dsRNA <221> Modified base <222> 1 <223> / Modified base = "Nucleotide: lacking 5'-phosphate group" <400> 130 ggagugugga uucgcacuc 19 <210> 131 <211> 19 <212> RNA <213> Artificial sequence <223> Description of artificial sequence: sense strand of dsRNA <221> Modified base <222> 1 <223> / Modified base = "Nucleotide: lacking 5'-phosphate group" <400> 131 cgggcgcacc ucucuuuac 19 <210> 132 <211> 19 <212> RNA <213> Artificial sequence <223> Description of artificial sequence: sense strand of dsRNA <221> Modified base <222> 1 <223> / Modified base = "Nucleotide: lacking 5'-phosphate group" <400> 132 gucucaaucg ccgcgucgc 19 <210> 133 <211> 19 <212> RNA <213> Artificial sequence <223> Description of artificial sequence: sense strand of dsRNA <221> Modified base <222> 1 <223> / Modified base = "Nucleotide: lacking 5'-phosphate group" <400> 133 auccauacug cggaacucc 19 <210> 134 <211> 19 <212> RNA <213> Artificial sequence <223> Artificial sequence description: Sense strand of dsRNA <221> Modified base <222> 1 <223> / Modified base = "Nucleotide: lacking 5'-phosphate group" <400> 134 cgcaccucuc uuuacgcgg 19 <210> 135 <211> 19 <212> RNA <213> Artificial sequence <223> Artificial sequence description: Sense strand of dsRNA <221> Modified base <222> 1 <223> / Modified base = "Nucleotide: lacking 5'-phosphate group" <400> 135 caacgaccga ccuugaggc 19 <210> 136 <211> 19 <212> RNA <213> Artificial sequence <223> Artificial sequence description: Sense strand of dsRNA <221> Modified base <222> 1 <223> / Modified base = "Nucleotide: lacking 5'-phosphate group" <400> 136 ccauacugcg gaacuccua 19 <210> 137 <211> 19 <212> RNA <213> Artificial sequence <223> Artificial sequence description: sense strand of dsRNA <221> Modified base <222> 1 <223> / Modified base = "Nucleotide: lacking 5'-phosphate group" <400> 137 ugaaucccgc ggacgaccc 19 <210> 138 <211> 19 <212> RNA <213> Artificial sequence <223> Artificial sequence description: sense strand of dsRNA <221> Modified base <222> 1 <223> / Modified base = "Nucleotide: lacking 5'-phosphate group" <400> 138 agaacucccu cgccucgca 19 <210> 139 <211> 19 <212> RNA <213> Artificial sequence <223> Artificial sequence description: sense strand of dsRNA <221> Modified base <222> 1 <223> / Modified base = "Nucleotide: lacking 5'-phosphate group" <400> 139 ggcgcaccuc ucuuuacgc 19 <210> 140 <211> 19 <212> RNA <213> Artificial sequence <223> Artificial sequence description: sense strand of dsRNA <221> Modified base <222> 1 <223> / Modified base = "Nucleotide: lacking 5'-phosphate group" <400> 140 gcgcaccucu cuuuacgcg 19 <210> 141 <211> 19 <212> RNA <213> Artificial sequence <223> Description of artificial sequence: sense strand of dsRNA <221> Modified base <222> 1 <223> / Modified base = "Nucleotide: lacking 5'-phosphate group" <400> 141 gcugaauccc gcggacgac 19 <210> 142 <211> 19 <212> RNA <213> Artificial sequence <223> Description of artificial sequence: sense strand of dsRNA <221> Modified base <222> 1 <223> / Modified base = "Nucleotide: lacking 5'-phosphate group" <400> 142 cacuucgcuu caccucugc 19 <210> 143 <211> 19 <212> RNA <213> Artificial sequence <223> Description of artificial sequence: sense strand of dsRNA <221> Modified base <222> 1 <223> / Modified base = "Nucleotide: lacking 5'-phosphate group" <400> 143 cucaaucgcc gcgucgcag 19 <210> 144 <211> 19 <212> RNA <213> Artificial sequence <223> Description of artificial sequence: sense strand of dsRNA <221> Modified base <222> 1 <223> / Modified base = "Nucleotide: lacking 5'-phosphate group" <400> 144 ucccgucggc gcugaaucc 19 <210> 145 <211> 19 <212> RNA <213> Artificial sequence <223> Description of artificial sequence: Sense strand of dsRNA <221> Modified base <222> 1 <223> / Modified base="Nucleotide: lacking 5'-phosphate group" <400> 145 cugaaucccg cggacgacc 19 <210> 146 <211> 19 <212> RNA <213> Artificial sequence <223> Description of artificial sequence: Sense strand of dsRNA <221> Modified base <222> 1 <223> / Modified base="Nucleotide: lacking 5'-phosphate group" <400> 146 agagucuaga cucguggug 19 <210> 147 <211> 19 <212> RNA <213> Artificial sequence <223> Description of artificial sequence: Sense strand of dsRNA <221> Modified base <222> 1 <223> / Modified base="Nucleotide: lacking 5'-phosphate group" <400> 147 uccauacugc ggaacuccu 19 <210> 148 <211> 19 <212> RNA <213> Artificial sequence <223> Description of artificial sequence: Sense strand of dsRNA <221> Modified base <222> 1 <223> / Modified base = "Nucleotide: lacking 5'-phosphate group" <400> 148 gcgcugaauc ccgcggacg 19 <210> 149 <211> 19 <212> RNA <213> Artificial sequence <223> Artificial sequence description: Sense strand of dsRNA <221> Modified base <222> 1 <223> / Modified base = "Nucleotide: lacking 5'-phosphate group" <400> 149 aguguggauu cgcacuccu 19 <210> 150 <211> 19 <212> RNA <213> Artificial sequence <223> Artificial sequence description: Sense strand of dsRNA <221> Modified base <222> 1 <223> / Modified base = "Nucleotide: lacking 5'-phosphate group" <400> 150 cccugcucgu guuacaggc 19 <210> 151 <211> 19 <212> RNA <213> Artificial sequence <223> Artificial sequence description: Sense strand of dsRNA <221> Modified base <222> 1 <223> / Modified base = "Nucleotide: lacking 5'-phosphate group" <400> 151 gaaucccgcg gacgacccg 19 <210> 152 <211> 19 <212> RNA <213> Artificial sequence <223> Artificial Sequence Description: Sense Strand of dsRNA <221> Modified Base <222> 1 <223> / Modified Base = "Nucleotide: Lacking 5'-phosphate group" <400> 152 aagcugugcc uuggguggc 19 <210> 153 <211> 19 <212> RNA <213> Artificial Sequence <223> Artificial Sequence Description: Sense Strand of dsRNA <221> Modified Base <222> 1 <223> / Modified Base = "Nucleotide: Lacking 5'-phosphate group" <400> 153 gcccugcucg uguuacagg 19 <210> 154 <211> 19 <212> RNA <213> Artificial Sequence <223> Artificial Sequence Description: Sense Strand of dsRNA <221> Modified Base <222> 1 <223> / Modified Base = "Nucleotide: Lacking 5'-phosphate group" <400> 154 gucccgucgg cgcugaauc 19 <210> 155 <211> 19 <212> RNA <213> Artificial Sequence <223> Artificial Sequence Description: Sense Strand of dsRNA <221> Modified Base <222> 1 <223> / Modified Base = "Nucleotide: Lacking 5'-phosphate group" <400> 155 aucuuaucaa cacuuccgg 19 <210> 156 <211> 19 <212> RNA <213> Artificial sequence <223> Description of artificial sequence: sense strand of dsRNA <221> Modified base <222> 1 <223> / Modified base = "Nucleotide: lacking 5'-phosphate group" <400> 156 cuuaucaaca cuuccggaa 19 <210> 157 <211> 19 <212> RNA <213> Artificial sequence <223> Description of artificial sequence: antisense strand of dsRNA <221> Modified base <222> 1 <223> / Modified base = "Nucleotide: lacking 5'-phosphate group" <400> 157 aaacgggcaa cauaccuug 19 <210> 158 <211> 19 <212> DNA <213> Artificial sequence <223> Description of artificial sequence: antisense strand of dsRNA <221> Modified base <222> 1 <223> / Modified base = "Nucleotide: lacking 5'-phosphate group" <221> Modified base <222> 2..19 <223> / Modified base = "Nucleotide corresponding to 2'-hydroxyl" <400> 158 taccaauuua ugccuacag 19 <210> 159 <211> 19 <212> RNA <213> Artificial sequence <223> Description of artificial sequence: antisense strand of dsRNA <221> Modified base <222> 1 <223> / Modified base = "Nucleotide: lacking 5'-phosphate group" <400> 159 uaugauaaaa cgccgcaga 19 <210> 160 <211> 19 <212> DNA <213> Artificial sequence <223> Artificial sequence description: Antisense strand of dsRNA <221> Modified base <222> 1 <223> / Modified base = "Nucleotide: lacking 5'-phosphate group" <221> Modified base <222> 2..19 <223> / Modified base = "Nucleotide corresponding to 2'-hydroxy" <400> 160 taugauaaaa cgccgcaga 19 <210> 161 <211> 19 <212> RNA <213> Artificial sequence <223> Artificial sequence description: Antisense strand of dsRNA <221> Modified base <222> 1 <223> / Modified base = "Nucleotide: lacking 5'-phosphate group" <400> 161 gagaugauua ggcagaggu 19 <210> 162 <211> 19 <212> DNA <213> Artificial sequence <223> Artificial sequence description: Antisense strand of dsRNA <221> Modified base <222> 1 <223> / Modified base = "Nucleotide: lacking 5'-phosphate group" <221> Modified base <222> 2..19 <223> / Modified base = "2'-hydroxy corresponding nucleotide" <400> 162 tacaaauggc acuaguaaa 19 <210> 163 <211> 19 <212> DNA <213> Artificial sequence <223> Artificial sequence description: Antisense strand of dsRNA <221> Modified base <222> 1 <223> / Modified base = "Nucleotide: lacking 5'-phosphate group" <221> Modified base <222> 2..19 <223> / Modified base = "2'-hydroxy corresponding nucleotide" <400> 163 tagaugauua ggcagaggu 19 <210> 164 <211> 19 <212> RNA <213> Artificial sequence <223> Artificial sequence description: Antisense strand of dsRNA <221> Modified base <222> 1 <223> / Modified base = "Nucleotide: lacking 5'-phosphate group" <400> 164 gaccaauuua ugccuacag 19 <210> 165 <211> 19 <212> RNA <213> Artificial sequence <223> Artificial sequence description: Antisense strand of dsRNA <221> Modified base <222> 1 <223> / Modified base = "Nucleotide: lacking 5'-phosphate group" <400> 165 ugauaaaacg ccgcagaca 19 <210> 166 <211> 19 <212> DNA <213> Artificial sequence <223> Description of artificial sequence: antisense strand of dsRNA <221> Modified base <222> 1 <223> / Modified base = "nucleotide: lacking 5'-phosphate group" <221> Modified base <222> 2..19 <223> / Modified base = "nucleotide corresponding to 2'-hydroxyl" <400> 166 tgauaaaacg ccgcagaca 19 <210> 167 <211> 19 <212> RNA <213> Artificial sequence <223> Description of artificial sequence: antisense strand of dsRNA <221> Modified base <222> 1 <223> / Modified base = "nucleotide: lacking 5'-phosphate group" <400> 167 ugaacaaaug gcacuagua 19 <210> 168 <211> 19 <212> DNA <213> Artificial sequence <223> Description of artificial sequence: antisense strand of dsRNA <221> Modified base <222> 1 <223> / Modified base = "nucleotide: lacking 5'-phosphate group" <221> Modified base <222> 2..19 <223> / Modified base = "nucleotide corresponding to 2'-hydroxyl" <400> 168 tgaacaaaug gcacuagua 19 <210> 169 <211> 19 <212> DNA <213> Artificial sequence <223> Description of artificial sequence: Antisense strand of dsRNA <221> Modified base <222> 1 <223> / Modified base = "Nucleotide: lacking 5'-phosphate group" <221> Modified base <222> 2..19 <223> / Modified base = "Nucleotide corresponding to 2'-hydroxy" <400> 169 tgcagaggug aaaaaguug 19 <210> 170 <211> 19 <212> RNA <213> Artificial sequence <223> Description of artificial sequence: Antisense strand of dsRNA <221> Modified base <222> 1 <223> / Modified base = "Nucleotide: lacking 5'-phosphate group" <400> 170 cgaaccacug aacaaaugg 19 <210> 171 <211> 19 <212> RNA <213> Artificial sequence <223> Description of artificial sequence: Antisense strand of dsRNA <221> Modified base <222> 1 <223> / Modified base = "Nucleotide: lacking 5'-phosphate group" <400> 171 ugcgucagca aacacuugg 19 <210> 172 <211> 19 <212> DNA <213> Artificial sequence <223> Description of artificial sequence: Antisense strand of dsRNA <221> Modified base <222> 1 <223> / Modified base = "Nucleotide: lacking 5'-phosphate group" <221> Modified base <222> 2..19 <223> / Modified base = "Nucleotide corresponding to 2'-hydroxyl" <400> 172 tgcgucagca aacacuugg 19 <210> 173 <211> 19 <212> DNA <213> Artificial sequence <223> Artificial sequence description: Antisense strand of dsRNA <221> Modified base <222> 1 <223> / Modified base = "Nucleotide: lacking 5'-phosphate group" <221> Modified base <222> 2..19 <223> / Modified base = "Nucleotide corresponding to 2'-hydroxyl" <400> 173 tgaaccacug aacaaaugg 19 <210> 174 <211> 19 <212> RNA <213> Artificial sequence <223> Artificial sequence description: Antisense strand of dsRNA <221> Modified base <222> 1 <223> / Modified base = "Nucleotide: lacking 5'-phosphate group" <400> 174 aacaaauggc acuaguaaa 19 <210> 175 <211> 19 <212> DNA <213> Artificial sequence <223> Artificial sequence description: Antisense strand of dsRNA <221> Modified base <222> 1 <223> / Modified base = "Nucleotide: lacking 5'-phosphate group" <221> Modified base <222> 2..19 <223> / Modified base = "Nucleotide corresponding to 2'-hydroxy" <400> 175 tgaugauuag gcagaggug 19 <210> 176 <211> 19 <212> RNA <213> Artificial sequence <223> Explanation of artificial sequence: Antisense strand of dsRNA <221> Modified base <222> 1 <223> / Modified base = "Nucleotide: lacking 5'-phosphate group" <400> 176 cacuaguaaa cugagccag 19 <210> 177 <211> 19 <212> DNA <213> Artificial sequence <223> Explanation of artificial sequence: Antisense strand of dsRNA <221> Modified base <222> 1 <223> / Modified base = "Nucleotide: lacking 5'-phosphate group" <221> Modified base <222> 2..19 <223> / Modified base = "Nucleotide corresponding to 2'-hydroxy" <400> 177 taacgggcaa cauaccuug 19 <210> 178 <211> 19 <212> DNA <213> Artificial sequence <223> Explanation of artificial sequence: Antisense strand of dsRNA <221> Modified base <222> 1 <223> / Modified base = "Nucleotide: lacking 5'-phosphate group" <221> Modified base <222> 2..19 <223> / Modified base = "2'-hydroxy corresponding nucleotide" <400> 178 tacuaguaaa cugagccag 19 <210> 179 <211> 19 <212> RNA <213> Artificial sequence <223> Artificial sequence description: Antisense strand of dsRNA <221> Modified base <222> 1 <223> / Modified base = "Nucleotide: lacking 5'-phosphate group" <400> 179 aauuuaugcc uacagccuc 19 <210> 180 <211> 19 <212> RNA <213> Artificial sequence <223> Artificial sequence description: Antisense strand of dsRNA <221> Modified base <222> 1 <223> / Modified base = "Nucleotide: lacking 5'-phosphate group" <400> 180 aaauggcacu aguaaacug 19 <210> 181 <211> 19 <212> RNA <213> Artificial sequence <223> Artificial sequence description: Antisense strand of dsRNA <221> Modified base <222> 1 <223> / Modified base = "Nucleotide: lacking 5'-phosphate group" <400> 181 acaaacgggc aacauaccu 19 <210> 182 <211> 19 <212> RNA <213> Artificial sequence <223> Description of artificial sequence: Antisense strand of dsRNA <221> Modified base <222> 1 <223> / Modified base = "Nucleotide: lacking 5'-phosphate group" <400> 182 uggacaaacg ggcaacaua 19 <210> 183 <211> 19 <212> DNA <213> Artificial sequence <223> Description of artificial sequence: Antisense strand of dsRNA <221> Modified base <222> 1 <223> / Modified base = "Nucleotide: lacking 5'-phosphate group" <221> Modified base <222> 2..19 <223> / Modified base = "2'-hydroxy corresponding nucleotide" <400> 183 tauuuaugcc uacagccuc 19 <210> 184 <211> 19 <212> RNA <213> Artificial sequence <223> Description of artificial sequence: Antisense strand of dsRNA <221> Modified base <222> 1 <223> / Modified base = "Nucleotide: lacking 5'-phosphate group" <400> 184 augauaaaac gccgcagac 19 <210> 185 <211> 19 <212> RNA <213> Artificial sequence <223> Description of artificial sequence: Antisense strand of dsRNA <221> Modified base <222> 1 <223> / Modified base = "Nucleotide: lacking 5'-phosphate group" <400> 185 ggcagaggug aaaaaguug 19 <210> 186 <211> 19 <212> RNA <213> Artificial sequence <223> Artificial sequence description: Antisense strand of dsRNA <221> Modified base <222> 1 <223> / Modified base = "Nucleotide: lacking 5'-phosphate group" <400> 186 ugaagcgaag ugcacacgg 19 <210> 187 <211> 19 <212> DNA <213> Artificial sequence <223> Artificial sequence description: Antisense strand of dsRNA <221> Modified base <222> 1 <223> / Modified base = "Nucleotide: lacking 5'-phosphate group" <221> Modified base <222> 2..19 <223> / Modified base = "2'-hydroxy corresponding nucleotide" <400> 187 tgaagcgaag ugcacacgg 19 <210> 188 <211> 19 <212> DNA <213> Artificial sequence <223> Artificial sequence description: Antisense strand of dsRNA <221> Modified base <222> 1 <223> / Modified base = "Nucleotide: lacking 5'-phosphate group" <221> Modified base <222> 2..19 <223> / Modified base = "2'-hydroxy corresponding nucleotide" <400> 188 tacgggcaac auaccuuga 19 <210> 189 <211> 19 <212> DNA <213> Artificial sequence <223> Artificial sequence description: Antisense strand of dsRNA <221> Modified base <222> 1 <223> / Modified base = "Nucleotide: lacking 5'-phosphate group" <221> Modified base <222> 2..19 <223> / Modified base = "2'-hydroxy corresponding nucleotide" <400> 189 taauggcacu aguaaacug 19 <210> 190 <211> 19 <212> RNA <213> Artificial sequence <223> Artificial sequence description: Antisense strand of dsRNA <221> Modified base <222> 1 <223> / Modified base = "Nucleotide: lacking 5'-phosphate group" <400> 190 aauugagaga aguccacca 19 <210> 191 <211> 19 <212> DNA <213> Artificial sequence <223> Artificial sequence description: Antisense strand of dsRNA <221> Modified base <222> 1 <223> / Modified base = "Nucleotide: lacking 5'-phosphate group" <221> Modified base <222> 2..19 <223> / Modified base = "2'-hydroxy corresponding nucleotide" <400> 191 tcaaacgggc aacauaccu 19 <210> 192 <211> 19 <212> RNA <213> Artificial sequence <223> Description of artificial sequence: Antisense strand of dsRNA <221> Modified base <222> 1 <223> / Modified base = "Nucleotide: lacking 5'-phosphate group" <400> 192 ccgccuguaa cacgagcag 19 <210> 193 <211> 19 <212> RNA <213> Artificial sequence <223> Description of artificial sequence: Antisense strand of dsRNA <221> Modified base <222> 1 <223> / Modified base = "Nucleotide: lacking 5'-phosphate group" <400> 193 aggacaaacg ggcaacaua 19 <210> 194 <211> 19 <212> RNA <213> Artificial sequence <223> Description of artificial sequence: Antisense strand of dsRNA <221> Modified base <222> 1 <223> / Modified base = "Nucleotide: lacking 5'-phosphate group" <400> 194 aacgggcaac auaccuuga 19 <210> 195 <211> 19 <212> RNA <213> Artificial sequence <223> Description of artificial sequence: Antisense strand of dsRNA <221> Modified base <222> 1 <223> / Modified base = "Nucleotide: lacking 5'-phosphate group" <400> 195 uccggaagug uugauaaga 19 <210> 196 <211> 19 <212> DNA <213> Artificial sequence <223> Artificial sequence description: Antisense strand of dsRNA <221> Modified base <222> 1 <223> / Modified base = "Nucleotide: lacking 5'-phosphate group" <221> Modified base <222> 2..19 <223> / Modified base = "Nucleotide corresponding to 2'-hydroxyl" <400> 196 tccggaagug uugauaaga 19 <210> 197 <211> 19 <212> RNA <213> Artificial sequence <223> Artificial sequence description: Antisense strand of dsRNA <221> Modified base <222> 1 <223> / Modified base = "Nucleotide: lacking 5'-phosphate group" <400> 197 agaugauuag gcagaggug 19 <210> 198 <211> 19 <212> RNA <213> Artificial sequence <223> Artificial sequence description: Antisense strand of dsRNA <221> Modified base <222> 1 <223> / Modified base = "Nucleotide: lacking 5'-phosphate group" <400> 198 aguccaagag uccucuuau 19 <210> 199 <211> 19 <212> DNA <213> Artificial sequence <223> Description of artificial sequence: Antisense strand of dsRNA <221> Modified base <222> 1 <223> / Modified base = "Nucleotide: lacking 5'-phosphate group" <221> Modified base <222> 2..19 <223> / Modified base = "Nucleotide corresponding to 2'-hydroxy" <400> 199 tugauaaaac gccgcagac 19 <210> 200 <211> 19 <212> RNA <213> Artificial sequence <223> Description of artificial sequence: Antisense strand of dsRNA <221> Modified base <222> 1 <223> / Modified base = "Nucleotide: lacking 5'-phosphate group" <400> 200 guccgcggga uucagcgcc 19 <210> 201 <211> 19 <212> RNA <213> Artificial sequence <223> Description of artificial sequence: Antisense strand of dsRNA <221> Modified base <222> 1 <223> / Modified base = "Nucleotide: lacking 5'-phosphate group" <400> 201 ugagaucuuc ugcgacgcg 19 <210> 202 <211> 19 <212> RNA <213> Artificial sequence <223> Artificial sequence description: Antisense strand of dsRNA <221> Modified base <222> 1 <223> / Modified base = "Nucleotide: lacking 5'-phosphate group" <400> 202 aaggucgguc guugacauu 19 <210> 203 <211> 19 <212> RNA <213> Artificial sequence <223> Artificial sequence description: Antisense strand of dsRNA <221> Modified base <222> 1 <223> / Modified base = "Nucleotide: lacking 5'-phosphate group" <400> 203 uggcacuagu aaacugagc 19 <210> 204 <211> 19 <212> DNA <213> Artificial sequence <223> Artificial sequence description: Antisense strand of dsRNA <221> Modified base <222> 1 <223> / Modified base = "Nucleotide: lacking 5'-phosphate group" <221> Modified base <222> 2..19 <223> / Modified base = "2'-hydroxy corresponding nucleotide" <400> 204 tauugagaga aguccacca 19 <210> 205 <211> 19 <212> RNA <213> Artificial sequence <223> Artificial sequence description: Antisense strand of dsRNA <221> Modified base <222> 1 <223> / Modified base = "Nucleotide: lacking 5'-phosphate group" <400> 205 ucuucugcga cgcggcgau 19 <210> 206 <211> 19 <212> RNA <213> Artificial sequence <223> Artificial sequence description: Antisense strand of dsRNA <221> Modified base <222> 1 <223> / Modified base = "Nucleotide: lacking 5'-phosphate group" <400> 206 gaaccacuga acaaauggc 19 <210> 207 <211> 19 <212> RNA <213> Artificial sequence <223> Artificial sequence description: Antisense strand of dsRNA <221> Modified base <222> 1 <223> / Modified base = "Nucleotide: lacking 5'-phosphate group" <400> 207 aguuccgcag uauggaucg 19 <210> 208 <211> 19 <212> RNA <213> Artificial sequence <223> Artificial sequence description: Antisense strand of dsRNA <221> Modified base <222> 1 <223> / Modified base = "Nucleotide: lacking 5'-phosphate group" <400> 208 gaugauuagg cagagguga 19 <210> 209 <211> 19 <212> RNA <213> Artificial sequence <223> Artificial sequence description: Antisense strand of dsRNA <221> Modified base <222> 1 <223> / Modified base = "Nucleotide: lacking 5'-phosphate group" <400> 209 aaaauugaga gaaguccac 19 <210> 210 <211> 19 <212> RNA <213> Artificial sequence <223> Artificial sequence description: Antisense strand of dsRNA <221> Modified base <222> 1 <223> / Modified base = "Nucleotide: lacking 5'-phosphate group" <400> 210 cccaagaaua uggugaccc 19 <210> 211 <211> 19 <212> RNA <213> Artificial sequence <223> Artificial sequence description: Antisense strand of dsRNA <221> Modified base <222> 1 <223> / Modified base = "Nucleotide: lacking 5'-phosphate group" <400> 211 agaucuucug cgacgcggc 19 <210> 212 <211> 19 <212> RNA <213> Artificial sequence <223> Artificial sequence description: Antisense strand of dsRNA <221> Modified base <222> 1 <223> / Modified base = "Nucleotide: lacking 5'-phosphate group" <400> 212 ucugcgacgc ggcgauuga 19 <210> 213 <211> 19 <212> RNA <213> Artificial sequence <223> Description of artificial sequence: Antisense strand of dsRNA <221> Modified base <222> 1 <223> / Modified base = "Nucleotide: lacking 5'-phosphate group" <400> 213 aacgccgcag acacaucca 19 <210> 214 <211> 19 <212> RNA <213> Artificial sequence <223> Description of artificial sequence: Antisense strand of dsRNA <221> Modified base <222> 1 <223> / Modified base = "Nucleotide: lacking 5'-phosphate group" <400> 214 cuuggaggcu ugaacagua 19 <210> 215 <211> 19 <212> RNA <213> Artificial sequence <223> Description of artificial sequence: Antisense strand of dsRNA <221> Modified base <222> 1 <223> / Modified base = "Nucleotide: lacking 5'-phosphate group" <400> 215 acaaauggca cuaguaaac 19 <210> 216 <211> 19 <212> RNA <213> Artificial sequence <223> Description of artificial sequence: Antisense strand of dsRNA <221> Modified base <222> 1 <223> / Modified base = "Nucleotide: lacking 5'-phosphate group" <400> 216 cugaacaaau ggcacuagu 19 <210> 217 <211> 19 <212> RNA <213> Artificial sequence <223> Description of artificial sequence: antisense strand of dsRNA <221> Modified base <222> 1 <223> / Modified base = "Nucleotide: lacking 5'-phosphate group" <400> 217 gagaucuucu gcgacgcgg 19 <210> 218 <211> 19 <212> RNA <213> Artificial sequence <223> Description of artificial sequence: antisense strand of dsRNA <221> Modified base <222> 1 <223> / Modified base = "Nucleotide: lacking 5'-phosphate group" <400> 218 cggaaguguu gauaagaua 19 <210> 219 <211> 19 <212> RNA <213> Artificial sequence <223> Description of artificial sequence: antisense strand of dsRNA <221> Modified base <222> 1 <223> / Modified base = "Nucleotide: lacking 5'-phosphate group" <400> 219 gggacugcga auuuuggcc 19 <210> 220 <211> 19 <212> RNA <213> Artificial sequence <223> Description of artificial sequence: antisense strand of dsRNA <221> Modified base <222> 1 <223> / Modified base = "Nucleotide: lacking 5'-phosphate group" <400> 220 augauuaggc agaggugaa 19 <210> 221 <211> 19 <212> RNA <213> Artificial sequence <223> Artificial sequence description: Antisense strand of dsRNA <221> Modified base <222> 1 <223> / Modified base = "Nucleotide: lacking 5'-phosphate group" <400> 221 auggcacuag uaaacugag 19 <210> 222 <211> 19 <212> RNA <213> Artificial sequence <223> Artificial sequence description: Antisense strand of dsRNA <221> Modified base <222> 1 <223> / Modified base = "Nucleotide: lacking 5'-phosphate group" <400> 222 agaggacaaa cgggcaaca 19 <210> 223 <211> 19 <212> RNA <213> Artificial sequence <223> Artificial sequence description: Antisense strand of dsRNA <221> Modified base <222> 1 <223> / Modified base = "Nucleotide: lacking 5'-phosphate group" <400> 223 cacugaacaa auggcacua 19 <210> 224 <211> 19 <212> RNA <213> Artificial sequence <223> Artificial sequence description: Antisense strand of dsRNA <221> Modified base <222> 1 <223> / Modified base = "Nucleotide: lacking 5'-phosphate group" <400> 224 caauuuaugc cuacagccu 19 <210> 225 <211> 19 <212> RNA <213> Artificial sequence <223> Artificial sequence description: Antisense strand of dsRNA <221> Modified base <222> 1 <223> / Modified base = "Nucleotide: lacking 5'-phosphate group" <400> 225 uaaaacgccg cagacacau 19 <210> 226 <211> 19 <212> DNA <213> Artificial sequence <223> Artificial sequence description: Antisense strand of dsRNA <221> Modified base <222> 1 <223> / Modified base = "Nucleotide: lacking 5'-phosphate group" <221> Modified base <222> 2..19 <223> / Modified base = "2'-hydroxyl corresponding nucleotide" <400> 226 taaaacgccg cagacacau 19 <210> 227 <211> 19 <212> RNA <213> Artificial sequence <223> Artificial sequence description: Antisense strand of dsRNA <221> Modified base <222> 1 <223> / Modified base = "Nucleotide: lacking 5'-phosphate group" <400> 227 ugcagaggug aagcgaagu 19 <210> 228 <211> 19 <212> RNA <213> Artificial sequence <223> Description of artificial sequence: Antisense strand of dsRNA <221> Modified base <222> 1 <223> / Modified base = "Nucleotide: lacking 5'-phosphate group" <400> 228 gugaagcgaa gugcacacg 19 <210> 229 <211> 19 <212> RNA <213> Artificial sequence <223> Description of artificial sequence: Antisense strand of dsRNA <221> Modified base <222> 1 <223> / Modified base = "Nucleotide: lacking 5'-phosphate group" <400> 229 auugagagaa guccaccac 19 <210> 230 <211> 19 <212> RNA <213> Artificial sequence <223> Description of artificial sequence: Antisense strand of dsRNA <221> Modified base <222> 1 <223> / Modified base = "Nucleotide: lacking 5'-phosphate group" <400> 230 auaaaacgcc gcagacaca 19 <210> 231 <211> 19 <212> RNA <213> Artificial sequence <223> Description of artificial sequence: Antisense strand of dsRNA <221> Modified base <222> 1 <223> / Modified base = "Nucleotide: lacking 5'-phosphate group" <400> 231 caaacgggca acauaccuu 19 <210> 232 <211> 19 <212> RNA <213> Artificial sequence <223> Artificial sequence description: Antisense strand of dsRNA <221> Modified base <222> 1 <223> / Modified base = "Nucleotide: lacking 5'-phosphate group" <400> 232 ccucaagguc ggucguuga 19 <210> 233 <211> 19 <212> RNA <213> Artificial sequence <223> Artificial sequence description: Antisense strand of dsRNA <221> Modified base <222> 1 <223> / Modified base = "Nucleotide: lacking 5'-phosphate group" <400> 233 guccaagagu ccucuuaug 19 <210> 234 <211> 19 <212> RNA <213> Artificial sequence <223> Artificial sequence description: Antisense strand of dsRNA <221> Modified base <222> 1 <223> / Modified base = "Nucleotide: lacking 5'-phosphate group" <400> 234 cucaaggucg gucguugac 19 <210> 235 <211> 19 <212> RNA <213> Artificial sequence <223> Description of artificial sequence: antisense strand of dsRNA <221> Modified base <222> 1 <223> / Modified base = "Nucleotide: lacking 5'-phosphate group" <400> 235 cucuuguucc caagaauau 19 <210> 236 <211> 19 <212> RNA <213> Artificial sequence <223> Description of artificial sequence: antisense strand of dsRNA <221> Modified base <222> 1 <223> / Modified base = "Nucleotide: lacking 5'-phosphate group" <400> 236 cccgccugua acacgagca 19 <210> 237 <211> 19 <212> RNA <213> Artificial sequence <223> Description of artificial sequence: antisense strand of dsRNA <221> Modified base <222> 1 <223> / Modified base = "Nucleotide: lacking 5'-phosphate group" <400> 237 uucugcgacg cggcgauug 19 <210> 238 <211> 19 <212> RNA <213> Artificial sequence <223> Description of artificial sequence: antisense strand of dsRNA <221> Modified base <222> 1 <223> / Modified base = "Nucleotide: lacking 5'-phosphate group" <400> 238 gcuuggaggc uugaacagu 19 <210> 239 <211> 19 <212> RNA <213> Artificial sequence <223> Description of artificial sequence: antisense strand of dsRNA <221> Modified base <222> 1 <223> / Modified base = "Nucleotide: lacking 5'-phosphate group" <400> 239 gaucuucugc gacgcggcg 19 <210> 240 <211> 19 <212> RNA <213> Artificial sequence <223> Description of artificial sequence: antisense strand of dsRNA <221> Modified base <222> 1 <223> / Modified base = "Nucleotide: lacking 5'-phosphate group" <400> 240 acgaaccacu gaacaaaug 19 <210> 241 <211> 19 <212> RNA <213> Artificial sequence <223> Description of artificial sequence: antisense strand of dsRNA <221> Modified base <222> 1 <223> / Modified base = "Nucleotide: lacking 5'-phosphate group" <400> 241 ucguccgcgg gauucagcg 19 <210> 242 <211> 19 <212> RNA <213> Artificial sequence <223> Description of artificial sequence: antisense strand of dsRNA <221> Modified base <222> 1 <223> / Modified base = "Nucleotide: lacking 5'-phosphate group" <400> 242 ccaagaauau ggugaccca 19 <210> 243 <211> 19 <212> RNA <213> Artificial sequence <223> Description of artificial sequence: Antisense strand of dsRNA <221> Modified base <222> 1 <223> / Modified base = "Nucleotide: lacking 5'-phosphate group" <400> 243 aguauggauc ggcagagga 19 <210> 244 <211> 19 <212> RNA <213> Artificial sequence <223> Description of artificial sequence: Antisense strand of dsRNA <221> Modified base <222> 1 <223> / Modified base = "Nucleotide: lacking 5'-phosphate group" <400> 244 caaggucggu cguugacau 19 <210> 245 <211> 19 <212> RNA <213> Artificial sequence <223> Description of artificial sequence: Antisense strand of dsRNA <221> Modified base <222> 1 <223> / Modified base = "Nucleotide: lacking 5'-phosphate group" <400> 245 ugagaugauu aggcagagg 19 <210> 246 <211> 19 <212> RNA <213> Artificial sequence <223> Description of artificial sequence: Antisense strand of dsRNA <221> Modified base <222> 1 <223> / Modified base = "Nucleotide: lacking 5'-phosphate group" <400> 246 gaagcgaagu gcacacggu 19 <210> 247 <211> 19 <212> RNA <213> Artificial sequence <223> Artificial sequence description: Antisense strand of dsRNA <221> Modified base <222> 1 <223> / Modified base = "Nucleotide: lacking 5'-phosphate group" <400> 247 uccgcaguau ggaucggca 19 <210> 248 <211> 19 <212> RNA <213> Artificial sequence <223> Artificial sequence description: Antisense strand of dsRNA <221> Modified base <222> 1 <223> / Modified base = "Nucleotide: lacking 5'-phosphate group" <400> 248 accacgaguc uagacucug 19 <210> 249 <211> 19 <212> RNA <213> Artificial sequence <223> Artificial sequence description: Antisense strand of dsRNA <221> Modified base <222> 1 <223> / Modified base = "Nucleotide: lacking 5'-phosphate group" <400> 249 agcuuggagg cuugaacag 19 <210> 250 <211> 19 <212> RNA <213> Artificial sequence <223> Description of artificial sequence: Antisense strand of dsRNA <221> Modified base <222> 1 <223> / Modified base = "Nucleotide: lacking 5'-phosphate group" <400> 250 auuuaugccu acagccucc 19 <210> 251 <211> 19 <212> RNA <213> Artificial sequence <223> Description of artificial sequence: Antisense strand of dsRNA <221> Modified base <222> 1 <223> / Modified base = "Nucleotide: lacking 5'-phosphate group" <400> 251 uuuaugccua cagccuccu 19 <210> 252 <211> 19 <212> RNA <213> Artificial sequence <223> Description of artificial sequence: Antisense strand of dsRNA <221> Modified base <222> 1 <223> / Modified base = "Nucleotide: lacking 5'-phosphate group" <400> 252 aaauugagag aaguccacc 19 <210> 253 <211> 19 <212> RNA <213> Artificial sequence <223> Description of artificial sequence: Antisense strand of dsRNA <221> Modified base <222> 1 <223> / Modified base = "Nucleotide: lacking 5'-phosphate group" <400> 253 gcagagguga aaaaguugc 19 <210> 254 <211> 19 <212> DNA <213> Artificial sequence <223> Description of artificial sequence: Antisense strand of dsRNA <221> Modified base <222> 1 <223> / Modified base = "Nucleotide: lacking 5'-phosphate group" <221> Modified base <222> 2..19 <223> / Modified base = "2'-hydroxy corresponding nucleotide" <400> 254 tcgccuguaa cacgagcag 19 <210> 255 <211> 19 <212> RNA <213> Artificial sequence <223> Description of artificial sequence: Antisense strand of dsRNA <221> Modified base <222> 1 <223> / Modified base = "Nucleotide: lacking 5'-phosphate group" <400> 255 acugaacaaa uggcacuag 19 <210> 256 <211> 19 <212> RNA <213> Artificial sequence <223> Description of artificial sequence: Antisense strand of dsRNA <221> Modified base <222> 1 <223> / Modified base = "Nucleotide: lacking 5'-phosphate group" <400> 256 ccgcaguaug gaucggcag 19 <210> 257 <211> 19 <212> RNA <213> Artificial sequence <223> Description of artificial sequence: Antisense strand of dsRNA <221> Modified base <222> 1 <223> / Modified base = "Nucleotide: lacking 5'-phosphate group" <400> 257 ggugaagcga agugcacac 19 <210> 258 <211> 19 <212> RNA <213> Artificial sequence <223> Artificial sequence description: Antisense strand of dsRNA <221> Modified base <222> 1 <223> / Modified base = "Nucleotide: lacking 5'-phosphate group" <400> 258 gcccgccugu aacacgagc 19 <210> 259 <211> 19 <212> RNA <213> Artificial sequence <223> Artificial sequence description: Antisense strand of dsRNA <221> Modified base <222> 1 <223> / Modified base = "Nucleotide: lacking 5'-phosphate group" <400> 259 gaaguguuga uaagauagg 19 <210> 260 <211> 19 <212> RNA <213> Artificial sequence <223> Artificial sequence description: Antisense strand of dsRNA <221> Modified base <222> 1 <223> / Modified base = "Nucleotide: lacking 5'-phosphate group" <400> 260 ugcgacgcgg cgauugaga 19 <210> 261 <211> 19 <212> RNA <213> Artificial sequence <223> Description of artificial sequence: Antisense strand of dsRNA <221> Modified base <222> 1 <223> / Modified base = "Nucleotide: lacking 5'-phosphate group" <400> 261 ugagaaggca cagacgggc 19 <210> 262 <211> 19 <212> RNA <213> Artificial sequence <223> Description of artificial sequence: Antisense strand of dsRNA <221> Modified base <222> 1 <223> / Modified base = "Nucleotide: lacking 5'-phosphate group" <400> 262 ggaaguguug auaagauag 19 <210> 263 <211> 19 <212> RNA <213> Artificial sequence <223> Description of artificial sequence: Antisense strand of dsRNA <221> Modified base <222> 1 <223> / Modified base = "Nucleotide: lacking 5'-phosphate group" <400> 263 gaggacaaac gggcaacau 19 <210> 264 <211> 19 <212> RNA <213> Artificial sequence <223> Description of artificial sequence: Antisense strand of dsRNA <221> Modified base <222> 1 <223> / Modified base = "Nucleotide: lacking 5'-phosphate group" <400> 264 ggacaaacgg gcaacauac 19 <210> 265 <211> 19 <212> RNA <213> Artificial Sequence <223> Artificial sequence description: antisense strand of dsRNA <221> Modified base <222> 1 <223> / Modified base = "Nucleotide: lacking 5'-phosphate group" <400> 265 gugcagaggu gaagcgaag 19 <210> 266 <211> 19 <212> RNA <213> Artificial Sequence <223> Artificial sequence description: antisense strand of dsRNA <221> Modified base <222> 1 <223> / Modified base = "Nucleotide: lacking 5'-phosphate group" <400> 266 aggugaagcg aagugcaca 19 <210> 267 <211> 19 <212> RNA <213> Artificial Sequence <223> Artificial sequence description: antisense strand of dsRNA <221> Modified base <222> 1 <223> / Modified base = "Nucleotide: lacking 5'-phosphate group" <400> 267 cgggacugcg aauuuuggc 19 <210> 268 <211> 19 <212> RNA <213> Artificial Sequence <223> Artificial sequence description: antisense strand of dsRNA <221> Modified base <222> 1 <223> / Modified base = "Nucleotide: lacking 5'-phosphate group" <400> 268 cgccuguaac acgagcagg 19 <210> 269 <211> 19 <212> RNA <213> Artificial sequence <223> Artificial sequence description: Antisense strand of dsRNA <221> Modified base <222> 1 <223> / Modified base = "Nucleotide: lacking 5'-phosphate group" <400> 269 agugcgaauc cacacucca 19 <210> 270 <211> 19 <212> RNA <213> Artificial sequence <223> Artificial sequence description: Antisense strand of dsRNA <221> Modified base <222> 1 <223> / Modified base = "Nucleotide: lacking 5'-phosphate group" <400> 270 ugccucaagg ucggucguu 19 <210> 271 <211> 19 <212> RNA <213> Artificial sequence <223> Artificial sequence description: Antisense strand of dsRNA <221> Modified base <222> 1 <223> / Modified base = "Nucleotide: lacking 5'-phosphate group" <400> 271 ccacgagucu agacucugu 19 <210> 272 <211> 19 <212> RNA <213> Artificial sequence <223> Artificial sequence description: antisense strand of dsRNA <221> Modified base <222> 1 <223> / Modified base = "Nucleotide: lacking 5'-phosphate group" <400> 272 cuucugcgac gcggcgauu 19 <210> 273 <211> 19 <212> RNA <213> Artificial sequence <223> Artificial sequence description: antisense strand of dsRNA <221> Modified base <222> 1 <223> / Modified base = "Nucleotide: lacking 5'-phosphate group" <400> 273 gacaaacggg caacauacc 19 <210> 274 <211> 19 <212> RNA <213> Artificial sequence <223> Artificial sequence description: antisense strand of dsRNA <221> Modified base <222> 1 <223> / Modified base = "Nucleotide: lacking 5'-phosphate group" <400> 274 uuccgcagua uggaucggc 19 <210> 275 <211> 19 <212> RNA <213> Artificial sequence <223> Artificial sequence description: antisense strand of dsRNA <221> Modified base <222> 1 <223> / Modified base = "Nucleotide: lacking 5'-phosphate group" <400> 275 aguguugaua agauagggc 19 <210> 276 <211> 19 <212> RNA <213> Artificial sequence <223> Description of artificial sequence: antisense strand of dsRNA <221> Modified base <222> 1 <223> / Modified base = "Nucleotide: lacking 5'-phosphate group" <400> 276 caaauggcac uaguaaacu 19 <210> 277 <211> 19 <212> RNA <213> Artificial sequence <223> Description of artificial sequence: antisense strand of dsRNA <221> Modified base <222> 1 <223> / Modified base = "Nucleotide: lacking 5'-phosphate group" <400> 277 ucaaggucgg ucguugaca 19 <210> 278 <211> 19 <212> RNA <213> Artificial sequence <223> Description of artificial sequence: antisense strand of dsRNA <221> Modified base <222> 1 <223> / Modified base = "Nucleotide: lacking 5'-phosphate group" <400> 278 cuagaaaauu gagagaagu 19 <210> 279 <211> 19 <212> RNA <213> Artificial sequence <223> Description of artificial sequence: antisense strand of dsRNA <221> Modified base <222> 1 <223> / Modified base = "Nucleotide: lacking 5'-phosphate group" <400> 279 gacaaaggac gucccgcgc 19 <210> 280 <211> 19 <212> RNA <213> Artificial sequence <223> Description of artificial sequence: Antisense strand of dsRNA <221> Modified base <222> 1 <223> / Modified base = "Nucleotide: lacking 5'-phosphate group" <400> 280 aguccaccac gagucuaga 19 <210> 281 <211> 19 <212> RNA <213> Artificial sequence <223> Description of artificial sequence: Antisense strand of dsRNA <221> Modified base <222> 1 <223> / Modified base = "Nucleotide: lacking 5'-phosphate group" <400> 281 gaguuccgca guauggauc 19 <210> 282 <211> 19 <212> RNA <213> Artificial sequence <223> Description of artificial sequence: Antisense strand of dsRNA <221> Modified base <222> 1 <223> / Modified base = "Nucleotide: lacking 5'-phosphate group" <400> 282 gcaguaugga ucggcagag 19 <210> 283 <211> 19 <212> RNA <213> Artificial sequence <223> Description of artificial sequence: Antisense strand of dsRNA <221> Modified base <222> 1 <223> / Modified base = "Nucleotide: lacking 5'-phosphate group" <400> 283 cgcaguaugg aucggcaga 19 <210> 284 <211> 19 <212> RNA <213> Artificial sequence <223> Artificial sequence description: Antisense strand of dsRNA <221> Modified base <222> 1 <223> / Modified base = "Nucleotide: lacking 5'-phosphate group" <400> 284 caguauggau cggcagagg 19 <210> 285 <211> 19 <212> RNA <213> Artificial sequence <223> Artificial sequence description: Antisense strand of dsRNA <221> Modified base <222> 1 <223> / Modified base = "Nucleotide: lacking 5'-phosphate group" <400> 285 accgcguaaa gagaggugc 19 <210> 286 <211> 19 <212> RNA <213> Artificial sequence <223> Artificial sequence description: Antisense strand of dsRNA <221> Modified base <222> 1 <223> / Modified base = "Nucleotide: lacking 5'-phosphate group" <400> 286 gcgaggcgag ggaguucuu 19 <210> 287 <211> 19 <212> RNA <213> Artificial sequence <223> Artificial sequence description: Antisense strand of dsRNA <221> Modified base <222> 1 <223> / Modified base = "Nucleotide: lacking 5'-phosphate group" <400> 287 cugcgaggcg agggaguuc 19 <210> 288 <211> 19 <212> RNA <213> Artificial sequence <223> Artificial sequence description: Antisense strand of dsRNA <221> Modified base <222> 1 <223> / Modified base = "Nucleotide: lacking 5'-phosphate group" <400> 288 gcccuagaaa auugagaga 19 <210> 289 <211> 19 <212> RNA <213> Artificial sequence <223> Artificial sequence description: Antisense strand of dsRNA <221> Modified base <222> 1 <223> / Modified base = "Nucleotide: lacking 5'-phosphate group" <400> 289 cguaaagaga ggugcgccc 19 <210> 290 <211> 19 <212> RNA <213> Artificial sequence <223> Artificial sequence description: Antisense strand of dsRNA <221> Modified base <222> 1 <223> / Modified base = "Nucleotide: lacking 5'-phosphate group" <400> 290 guuccgcagu auggaucgg 19 <210> 291 <211> 19 <212> RNA <213> Artificial sequence <223> Description of artificial sequence: Antisense strand of dsRNA <221> Modified base <222> 1 <223> / Modified base = "Nucleotide: lacking 5'-phosphate group" <400> 291 ucugcgaggc gagggaguu 19 <210> 292 <211> 19 <212> RNA <213> Artificial sequence <223> Description of artificial sequence: Antisense strand of dsRNA <221> Modified base <222> 1 <223> / Modified base = "Nucleotide: lacking 5'-phosphate group" <400> 292 guauggaucg gcagaggag 19 <210> 293 <211> 19 <212> RNA <213> Artificial sequence <223> Description of artificial sequence: Antisense strand of dsRNA <221> Modified base <222> 1 <223> / Modified base = "Nucleotide: lacking 5'-phosphate group" <400> 293 gagugcgaau ccacacucc 19 <210> 294 <211> 19 <212> RNA <213> Artificial sequence <223> Description of artificial sequence: Antisense strand of dsRNA <221> Modified base <222> 1 <223> / Modified base = "Nucleotide: lacking 5'-phosphate group" <400> 294 guaaagagag gugcgcccg 19 <210> 295 <211> 19 <212> RNA <213> Artificial sequence <223> Description of artificial sequence: Antisense strand of dsRNA <221> Modified base <222> 1 <223> / Modified base = "Nucleotide: lacking 5'-phosphate group" <400> 295 gcgacgcggc gauugagac 19 <210> 296 <211> 19 <212> RNA <213> Artificial sequence <223> Description of artificial sequence: Antisense strand of dsRNA <221> Modified base <222> 1 <223> / Modified base = "Nucleotide: lacking 5'-phosphate group" <400> 296 ggaguuccgc aguauggau 19 <210> 297 <211> 19 <212> RNA <213> Artificial sequence <223> Description of artificial sequence: Antisense strand of dsRNA <221> Modified base <222> 1 <223> / Modified base = "Nucleotide: lacking 5'-phosphate group" <400> 297 ccgcguaaag agaggugcg 19 <210> 298 <211> 19 <212> RNA <213> Artificial sequence <223> Description of artificial sequence: Antisense strand of dsRNA <221> Modified base <222> 1 <223> / Modified base = "Nucleotide: lacking 5'-phosphate group" <400> 298 gccucaaggu cggucguug 19 <210> 299 <211> 19 <212> RNA <213> Artificial sequence <223> Artificial sequence description: Antisense strand of dsRNA <221> Modified base <222> 1 <223> / Modified base = "Nucleotide: lacking 5'-phosphate group" <400> 299 uaggaguucc gcaguaugg 19 <210> 300 <211> 19 <212> RNA <213> Artificial sequence <223> Artificial sequence description: Antisense strand of dsRNA <221> Modified base <222> 1 <223> / Modified base = "Nucleotide: lacking 5'-phosphate group" <400> 300 gggucguccg cgggauuca 19 <210> 301 <211> 19 <212> RNA <213> Artificial sequence <223> Artificial sequence description: Antisense strand of dsRNA <221> Modified base <222> 1 <223> / Modified base = "Nucleotide: lacking 5'-phosphate group" <400> 301 ugcgaggcga gggaguucu 19 <210> 302 <211> 19 <212> RNA <213> Artificial sequence <223> Artificial Sequence Description: Antisense Strand of dsRNA <221> Modified Base <222> 1 <223> / Modified Base = "Nucleotide: Lacking 5'-Phosphate Group" <400> 302 gcguaaagag aggugcgcc 19 <210> 303 <211> 19 <212> RNA <213> Artificial Sequence <223> Artificial Sequence Description: Antisense Strand of dsRNA <221> Modified Base <222> 1 <223> / Modified Base = "Nucleotide: Lacking 5'-Phosphate Group" <400> 303 cgcguaaaga gaggugcgc 19 <210> 304 <211> 19 <212> RNA <213> Artificial Sequence <223> Artificial Sequence Description: Antisense Strand of dsRNA <221> Modified Base <222> 1 <223> / Modified Base = "Nucleotide: Lacking 5'-Phosphate Group" <400> 304 gucguccgcg ggauucagc 19 <210> 305 <211> 19 <212> RNA <213> Artificial Sequence <223> Artificial Sequence Description: Antisense Strand of dsRNA <221> Modified Base <222> 1 <223> / Modified Base = "Nucleotide: Lacking 5'-Phosphate Group" <400> 305 gcagagguga agcgaagug 19 <210> 306 <211> 19 <212> RNA <213> Artificial sequence <223> Artificial sequence description: Antisense strand of dsRNA <221> Modified base <222> 1 <223> / Modified base = "Nucleotide: lacking 5'-phosphate group" <400> 306 cugcgacgcg gcgauugag 19 <210> 307 <211> 19 <212> RNA <213> Artificial sequence <223> Artificial sequence description: Antisense strand of dsRNA <221> Modified base <222> 1 <223> / Modified base = "Nucleotide: lacking 5'-phosphate group" <400> 307 ggauucagcg ccgacggga 19 <210> 308 <211> 19 <212> RNA <213> Artificial sequence <223> Artificial sequence description: Antisense strand of dsRNA <221> Modified base <222> 1 <223> / Modified base = "Nucleotide: lacking 5'-phosphate group" <400> 308 ggucguccgc gggauucag 19 <210> 309 <211> 19 <212> RNA <213> Artificial sequence <223> Artificial sequence description: Antisense strand of dsRNA <221> Modified base <222> 1 <223> / Modified base = "Nucleotide: lacking 5'-phosphate group" <400> 309 caccacgagu cuagacucu 19 <210> 310 <211> 19 <212> RNA <213> Artificial sequence <223> Explanation of artificial sequence: antisense strand of dsRNA <221> Modified base <222> 1 <223> / Modified base = "Nucleotide: lacking 5'-phosphate group" <400> 310 aggaguuccg caguaugga 19 <210> 311 <211> 19 <212> RNA <213> Artificial sequence <223> Explanation of artificial sequence: antisense strand of dsRNA <221> Modified base <222> 1 <223> / Modified base = "Nucleotide: lacking 5'-phosphate group" <400> 311 cguccgcggg auucagcgc 19 <210> 312 <211> 19 <212> RNA <213> Artificial sequence <223> Explanation of artificial sequence: antisense strand of dsRNA <221> Modified base <222> 1 <223> / Modified base = "Nucleotide: lacking 5'-phosphate group" <400> 312 aggagugcga auccacacu 19 <210> 313 <211> 19 <212> RNA <213> Artificial sequence <223> Explanation of artificial sequence: antisense strand of dsRNA <221> Modified base <222> 1 <223> / Modified base = "Nucleotide: lacking 5'-phosphate group" <400> 313 gccuguaaca cgagcaggg 19 <210> 314 <211> 19 <212> RNA <213> Artificial sequence <223> Artificial sequence description: Antisense strand of dsRNA <221> Modified base <222> 1 <223> / Modified base = "Nucleotide: lacking 5'-phosphate group" <400> 314 cgggucgucc gcgggauuc 19 <210> 315 <211> 19 <212> RNA <213> Artificial sequence <223> Artificial sequence description: Antisense strand of dsRNA <221> Modified base <222> 1 <223> / Modified base = "Nucleotide: lacking 5'-phosphate group" <400> 315 gccacccaag gcacagcuu 19 <210> 316 <211> 19 <212> RNA <213> Artificial sequence <223> Artificial sequence description: Antisense strand of dsRNA <221> Modified base <222> 1 <223> / Modified base = "Nucleotide: lacking 5'-phosphate group" <400> 316 ccuguaacac gagcagggc 19 <210> 317 <211> 19 <212> RNA <213> Artificial sequence <223> Artificial sequence description: Antisense strand of dsRNA <221> Modified base <222> 1 <223> / Modified base = "Nucleotide: lacking 5'-phosphate group" <400> 317 gauucagcgc cgacgggac 19 <210> 318 <211> 19 <212> RNA <213> Artificial sequence <223> Artificial sequence description: Antisense strand of dsRNA <221> Modified base <222> 1 <223> / Modified base = "Nucleotide: lacking 5'-phosphate group" <400> 318 ccggaagugu ugauaagau 19 <210> 319 <211> 19 <212> RNA <213> Artificial sequence <223> Artificial sequence description: Antisense strand of dsRNA <221> Modified base <222> 1 <223> / Modified base = "Nucleotide: lacking 5'-phosphate group" <400> 319 uuccggaagu guugauaag 19 <210> 320 <211> 19 <212> DNA <213> Artificial sequence <223> Artificial sequence description: Antisense strand of dsRNA <221> Modified base <222> 1 <223> / Modified base = "Nucleotide: lacking 5'-phosphate group" <221> Modified base <222> 2..19 <223> / Modified base = "2'-hydroxyl corresponding nucleotide" <400> 320 tuccggaagu guugauaag 19 <210> 321 <211> 21 <212> DNA <213> Artificial sequence <223> Description of artificial sequence: sense strand of dsRNA <221> Modified base <222> 1 <223> / Modified base = "nucleotide: lacking 5'-phosphate group" <221> Modified base <222> 1, 2, 6, 8, 10, 11, 13, 14, 15, 17, 18, 19 <223> / Modified base = "2'-O-methyl corresponding nucleotide" <221> Modified base <222> 21 <223> / Modified base = "5'-thio-phosphate thymidine" <221> Modified base <222> 3, 4, 5, 7, 9, 12, 16 <223> / Modified base = "2'-hydroxy corresponding nucleotide" <400> 321 caagguaugu ugcccguuut t 21 <210> 322 <211> 20 <212> DNA <213> Artificial sequence <223> Description of artificial sequence: sense strand of dsRNA <221> Modified base <222> 1 <223> / Modified base = "nucleotide: lacking 5'-phosphate group" <221> Modified base <222> 2, 4, 6, 8, 10, 12, 14, 16, 18 <223> / Modified base = "2'-O-methyl corresponding nucleotide" <221> Modified base <222> 1, 3, 5, 7, 9, 11, 13, 15, 17, 19 <223> / Modified base = "2'-deoxy-2'-fluoro corresponding nucleotide" <221> Modified base <222> 20 <223> / Modified base = "3'-3'-linked deoxythymidine" <400> 322 cuguaggcau aaauugguat 20 <210> 323 <211> 21 <212> DNA <213> Artificial sequence <223> Artificial sequence description: Sense strand of dsRNA <221> Modified base <222> 1 <223> / Modified base = "Nucleotide: lacking 5'-phosphate group" <221> Modified base <222> 1, 2, 3, 5, 8, 10, 11, 12, 13, 15, 16, 18 <223> / Modified base = "2'-O-methyl corresponding nucleotide" <221> Modified base <222> 21 <223> / Modified base = "5'-thiophosphate thymidine" <221> Modified base <222> 4, 6, 7, 9, 14, 17, 19 <223> / Modified base = "2'-hydroxy corresponding nucleotide" <400> 323 ucugcggcgu uuuaucauat t 21 <210> 324 <211> 20 <212> DNA <213> Artificial sequence <223> Artificial sequence description: Sense strand of dsRNA <221> Modified base <222> 1 <223> / Modified base = "Nucleotide: lacking 5'-phosphate group" <221> Modified base <222> 2, 4, 6, 8, 10, 12, 14, 16, 18 <223> / Modified base = "2'-O-methyl corresponding nucleotide" <221> Modified base <222> 1, 3, 5, 7, 9, 11, 13, 15, 17, 19 <223> / Modified base = "2'-deoxy-2'-fluoro corresponding nucleotide" <221> Modified base <222> 20 <223> / Modified base = "3'-3'-linked deoxythymidine" <400> 324 ucugcggcgu uuuaucauat 20 <210> 325 <211> 21 <212> DNA <213> Artificial sequence <223> Artificial sequence description: sense strand of dsRNA <221> Modified base <222> 1 <223> / Modified base = "Nucleotide: lacking 5'-phosphate group" <221> Modified base <222> 1, 2, 3, 4, 5, 6, 8, 9, 10, 13, 14, 16, 17, 18, 19 <223> / Modified base = "2'-O-methyl corresponding nucleotide" <221> Modified base <222> 21 <223> / Modified base = "5'-thiophosphate thymidine" <221> Modified base <222> 7, 11, 12, 15 <223> / Modified base = "2'-hydroxy corresponding nucleotide" <400> 325 accucugccu aaucaucuct t 21 <210> 326 <211> 20 <212> DNA <213> Artificial sequence <223> Description of artificial sequence: sense strand of dsRNA <221> Modified base <222> 1 <223> / Modified base = "Nucleotide: lacking 5'-phosphate group" <221> Modified base <222> 2, 4, 6, 8, 10, 12, 14, 16, 18 <223> / Modified base = "2'-O-methyl corresponding nucleotide" <221> Modified base <222> 1, 3, 5, 7, 9, 11, 13, 15, 17, 19 <223> / Modified base = "2'-deoxy-2'-fluoro corresponding nucleotide" <221> Modified base <222> 20 <223> / Modified base = "3'-3'-linked deoxythymidine" <400> 326 uuuacuagug ccauuuguat 20 <210> 327 <211> 20 <212> DNA <213> Artificial sequence <223> Description of artificial sequence: sense strand of dsRNA <221> Modified base <222> 1 <223> / Modified base = "Nucleotide: lacking 5'-phosphate group" <221> Modified base <222> 2, 4, 6, 8, 10, 12, 14, 16, 18 <223> / Modified base = "2'-O-methyl corresponding nucleotide" <221> Modified base <222> 1, 3, 5, 7, 9, 11, 13, 15, 17, 19 <223> / Modified base = "2'-deoxy-2'-fluoro corresponding nucleotide" <221> Modified base <222> 20 <223> / Modified base = "3'-3'-linked deoxythymidine" <400> 327 accucugccu aaucaucuat 20 <210> 328 <211> 21 <212> DNA <213> Artificial sequence <223> Artificial sequence description: Sense strand of dsRNA <221> Modified base <222> 1 <223> / Modified base = "Nucleotide: lacking 5'-phosphate group" <221> Modified base <222> 1, 2, 4, 8, 10, 14, 15, 18, 19 <223> / Modified base = "2'-O-methyl corresponding nucleotide" <221> Modified base <222> 21 <223> / Modified base = "5'-thiophosphate thymidine" <221> Modified base <222> 3, 5, 6, 7, 9, 11, 12, 13, 16, 17 <223> / Modified base = "2'-hydroxy corresponding nucleotide" <400> 328 cuguaggcau aaauugguct t 21 <210> 329 <211> 21 <212> DNA <213> Artificial sequence <223> Artificial sequence description: Sense strand of dsRNA <221> Modified base <222> 1 <223> / Modified base = "Nucleotide: lacking 5'-phosphate group" <221> Modified base <222> 1, 2, 3, 4, 5, 7, 10, 12, 13, 14, 15, 17, 18 <223> / Modified base = "2'-O-methyl corresponding nucleotide" <221> Modified base <222> 21 <223> / Modified base = "5'-thio-phosphate thymidine" <221> Modified base <222> 6, 8, 9, 11, 16, 19 <223> / Modified base = "2'-hydroxy corresponding nucleotide" <400> 329 ugucugcggc guuuuaucat t 21 <210> 330 <211> 20 <212> DNA <213> Artificial sequence <223> Artificial sequence description: sense strand of dsRNA <221> Modified base <222> 1 <223> / Modified base = "Nucleotide: lacking 5'-phosphate group" <221> Modified base <222> 2, 4, 6, 8, 10, 12, 14, 16, 18 <223> / Modified base = "2'-O-methyl corresponding nucleotide" <221> Modified base <222> 1, 3, 5, 7, 9, 11, 13, 15, 17, 19 <223> / Modified base = "2'-deoxy-2'-fluoro corresponding nucleotide" <221> Modified base <222> 20 <223> / Modified base = "3'-3'-linked deoxythymidine" <400> 330 ugucugcggc guuuuaucat 20 <210> 331 <211> 21 <212> DNA <213> Artificial sequence <223> Description of artificial sequence: Sense strand of dsRNA <221> Modified base <222> 1 <223> / Modified base = "Nucleotide: lacking 5'-phosphate group" <221> Modified base <222> 1, 2, 3, 4, 7, 9, 10, 12, 13, 14, 16, 17, 18 <223> / Modified base = "2'-O-methyl corresponding nucleotide" <221> Modified base <222> 21 <223> / Modified base = "5'-thio-phosphate thymidine" <221> Modified base <222> 5, 6, 8, 11, 15, 19 <223> / Modified base = "2'-hydroxy corresponding nucleotide" <400> 331 uacuagugcc auuuguucat t 21 <210> 332 <211> 20 <212> DNA <213> Artificial sequence <223> Description of artificial sequence: Sense strand of dsRNA <221> Modified base <222> 1 <223> / Modified base = "Nucleotide: lacking 5'-phosphate group" <221> Modified base <222> 2, 4, 6, 8, 10, 12, 14, 16, 18 <223> / Modified base = "2'-O-methyl corresponding nucleotide" <221> Modified base <222> 1, 3, 5, 7, 9, 11, 13, 15, 17, 19 <223> / Modified base = "2'-deoxy-2'-fluoro corresponding nucleotide" <221> Modified base <222> 20 <223> / Modified base = "3'-3'-linked deoxythymidine" <400> 332 uacuagugcc auuuguucat 20 <210> 333 <211> 20 <212> DNA <213> Artificial sequence <223> Artificial sequence description: Sense strand of dsRNA <221> Modified base <222> 1 <223> / Modified base = "Nucleotide: lacking 5'-phosphate group" <221> Modified base <222> 2, 4, 6, 8, 10, 12, 14, 16, 18 <223> / Modified base = "2'-O-methyl corresponding nucleotide" <221> Modified base <222> 1, 3, 5, 7, 9, 11, 13, 15, 17, 19 <223> / Modified base = "2'-deoxy-2'-fluoro corresponding nucleotide" <221> Modified base <222> 20 <223> / Modified base = "3'-3'-linked deoxythymidine" <400> 333 caacuuuuuc accucugcat 20 <210> 334 <211> 21 <212> DNA <213> Artificial sequence <223> Artificial sequence description: Sense strand of dsRNA <221> Modified base <222> 1 <223> / Modified base = "Nucleotide: lacking 5'-phosphate group" <221> Modified base <222> 1, 2, 4, 5, 6, 8, 9, 10, 13, 16, 17, 18 <223> / Modified base = "2'-O-methyl corresponding nucleotide" <221> Modified base <222> 21 <223> / Modified base = "5'-thiophosphate thymidine" <221> Modified base <222> 3, 7, 11, 12, 14, 15, 19 <223> / Modified base = "2'-hydroxy corresponding nucleotide" <400> 334 ccauuuguuc agugguucgt t 21 <210> 335 <211> 21 <212> DNA <213> Artificial sequence <223> Artificial sequence description: Sense strand of dsRNA <221> Modified base <222> 1 <223> / Modified base = "Nucleotide: lacking 5'-phosphate group" <221> Modified base <222> 1, 2, 6, 8, 9, 10, 12, 13, 16, 18 <223> / Modified base = "2'-O-methyl corresponding nucleotide" <221> Modified base <222> 21 <223> / Modified base = "5'-thiophosphate thymidine" <221> Modified base <222> 3, 4, 5, 7, 11, 14, 15, 17, 19 <223> / Modified base = "2'-hydroxy corresponding nucleotide" <400> 335 ccaaguguuu gcugacgcat t 21 <210> 336 <211> 20 <212> DNA <213> Artificial sequence <223> Artificial sequence description: Sense strand of dsRNA <221> Modified base <222> 1 <223> / Modified base = "Nucleotide: lacking 5'-phosphate group" <221> Modified base <222> 2, 4, 6, 8, 10, 12, 14, 16, 18 <223> / Modified base = "2'-O-methyl corresponding nucleotide" <221> Modified base <222> 1, 3, 5, 7, 9, 11, 13, 15, 17, 19 <223> / Modified base = "2'-deoxy-2'-fluoro corresponding nucleotide" <221> Modified base <222> 20 <223> / Modified base = "3'-3'-linked deoxythymidine" <400> 336 ccaaguguuu gcugacgcat 20 <210> 337 <211> 20 <212> DNA <213> Artificial sequence <223> Description of artificial sequence: sense strand of dsRNA <221> Modified base <222> 1 <223> / Modified base = "Nucleotide: lacking 5'-phosphate group" <221> Modified base <222> 2, 4, 6, 8, 10, 12, 14, 16, 18 <223> / Modified base = "2'-O-methyl corresponding nucleotide" <221> Modified base <222> 1, 3, 5, 7, 9, 11, 13, 15, 17, 19 <223> / Modified base = "2'-deoxy-2'-fluoro corresponding nucleotide" <221> Modified base <222> 20 <223> / Modified base = "3'-3'-linked deoxythymidine" <400> 337 ccauuuguuc agugguucat 20 <210> 338 <211> 21 <212> DNA <213> Artificial sequence <223> Description of artificial sequence: sense strand of dsRNA <221> Modified base <222> 1 <223> / Modified base = "nucleotide: lacking 5'-phosphate group" <221> Modified base <222> 1, 2, 3, 5, 6, 9, 11, 12, 14, 15, 16, 18, 19 <223> / Modified base = "2'-O-methyl corresponding nucleotide" <221> Modified base <222> 21 <223> / Modified base = "5'-thio-phosphate thymidine" <221> Modified base <222> 4, 7, 8, 10, 13, 17 <223> / Modified base = "2'-hydroxy corresponding nucleotide" <400> 338 uuuacuagug ccauuuguut t 21 <210> 339 <211> 20 <212> DNA <213> Artificial sequence <223> Description of artificial sequence: sense strand of dsRNA <221> Modified base <222> 1 <223> / Modified base = "nucleotide: lacking 5'-phosphate group" <221> Modified base <222> 2, 4, 6, 8, 10, 12, 14, 16, 18 <223> / Modified base = "2'-O-methyl corresponding nucleotide" <221> Modified base <222> 1, 3, 5, 7, 9, 11, 13, 15, 17, 19 <223> / Modified base = "2'-deoxy-2'-fluoro corresponding nucleotide" <221> Modified base <222> 20 <223> / Modified base = "3'-3'-linked deoxythymidine" <400> 339 caccucugcc uaaucaucat 20 <210> 340 <211> 21 <212> DNA <213> Artificial sequence <223> Artificial sequence description: Sense strand of dsRNA <221> Modified base <222> 1 <223> / Modified base = "Nucleotide: lacking 5'-phosphate group" <221> Modified base <222> 1, 2, 5, 6, 7, 10, 11, 12, 14, 15, 18 <223> / Modified base = "2'-O-methyl corresponding nucleotide" <221> Modified base <222> 21 <223> / Modified base = "5'-thiophosphate thymidine" <221> Modified base <222> 3, 4, 8, 9, 13, 16, 17, 19 <223> / Modified base = "2'-hydroxy corresponding nucleotide" <400> 340 cuggcucagu uuacuagugt t 21 <210> 341 <211> 20 <212> DNA <213> Artificial sequence <223> Artificial sequence description: Sense strand of dsRNA <221> Modified base <222> 1 <223> / Modified base = "Nucleotide: lacking 5'-phosphate group" <221> Modified base <222> 2, 4, 6, 8, 10, 12, 14, 16, 18 <223> / Modified base = "2'-O-methyl corresponding nucleotide" <221> Modified base <222> 1, 3, 5, 7, 9, 11, 13, 15, 17, 19 <223> / Modified base = "2'-deoxy-2'-fluoro corresponding nucleotide" <221> Modified base <222> 20 <223> / Modified base = "3'-3'-linked deoxythymidine" <400> 341 caagguaugu ugcccguuat 20 <210> 342 <211> 20 <212> DNA <213> Artificial sequence <223> Artificial sequence description: sense strand of dsRNA <221> Modified base <222> 1 <223> / Modified base = "Nucleotide: lacking 5'-phosphate group" <221> Modified base <222> 2, 4, 6, 8, 10, 12, 14, 16, 18 <223> / Modified base = "2'-O-methyl corresponding nucleotide" <221> Modified base <222> 1, 3, 5, 7, 9, 11, 13, 15, 17, 19 <223> / Modified base = "2'-deoxy-2'-fluoro corresponding nucleotide" <221> Modified base <222> 20 <223> / Modified base = "3'-3'-linked deoxythymidine" <400> 342 cuggcucagu uuacuaguat 20 <210> 343 <211> 21 <212> DNA <213> Artificial sequence <223> Description of artificial sequence: sense strand of dsRNA <221> Modified base <222> 1 <223> / Modified base = "Nucleotide: lacking 5'-phosphate group" <221> Modified base <222> 1, 2, 5, 6, 8, 12, 14, 18, 19 <223> / Modified base = "2'-O-methyl corresponding nucleotide" <221> Modified base <222> 21 <223> / Modified base = "5'-thio-phosphate thymidine" <221> Modified base <222> 3, 4, 7, 9, 10, 11, 13, 15, 16, 17 <223> / Modified base = "2'-hydroxy corresponding nucleotide" <400> 343 gaggcuguag gcauaaauut t 21 <210> 344 <211> 21 <212> DNA <213> Artificial sequence <223> Description of artificial sequence: sense strand of dsRNA <221> Modified base <222> 1 <223> / Modified base = "Nucleotide: lacking 5'-phosphate group" <221> Modified base <222> 1, 2, 4, 5, 6, 8, 9, 12, 14, 15, 17, 18, 19 <223> / Modified base = "2'-O-methyl corresponding nucleotide" <221> Modified base <222> 21 <223> / Modified base = "5'-thio-phosphate thymidine" <221> Modified base <222> 3, 7, 10, 11, 13, 16 <223> / Modified base = "2'-hydroxy corresponding nucleotide" <400> 344 caguuuacua gugccauuut t 21 <210> 345 <211> 21 <212> DNA <213> Artificial sequence <223> Artificial sequence description: Sense strand of dsRNA <221> Modified base <222> 1 <223> / Modified base = "Nucleotide: lacking 5'-phosphate group" <221> Modified base <222> 1, 2, 4, 6, 8, 9, 11, 12, 13, 15, 16, 17, 19 <223> / Modified base = "2'-O-methyl corresponding nucleotide" <221> Modified base <222> 21 <223> / Modified base = "5'-thiophosphate thymidine" <221> Modified base <222> 3, 5, 7, 10, 14, 18 <223> / Modified base = "2'-hydroxy corresponding nucleotide" <400> 345 agguauguug cccguuugut t 21 <210> 346 <211> 20 <212> DNA <213> Artificial sequence <223> Artificial sequence description: Sense strand of dsRNA <221> Modified base <222> 1 <223> / Modified base = "Nucleotide: lacking 5'-phosphate group" <221> Modified base <222> 2, 4, 6, 8, 10, 12, 14, 16, 18 <223> / Modified base = "2'-O-methyl corresponding nucleotide" <221> Modified base <222> 1, 3, 5, 7, 9, 11, 13, 15, 17, 19 <223> / Modified base = "2'-deoxy-2'-fluoro corresponding nucleotide" <221> Modified base <222> 20 <223> / Modified base = "3'-3'-linked deoxythymidine" <400> 346 uauguugccc guuuguccat 20 <210> 347 <211> 20 <212> DNA <213> Artificial sequence <223> Artificial sequence description: Sense strand of dsRNA <221> Modified base <222> 1 <223> / Modified base = "Nucleotide: lacking 5'-phosphate group" <221> Modified base <222> 2, 4, 6, 8, 10, 12, 14, 16, 18 <223> / Modified base = "2'-O-methyl corresponding nucleotide" <221> Modified base <222> 1, 3, 5, 7, 9, 11, 13, 15, 17, 19 <223> / Modified base = "2'-deoxy-2'-fluoro corresponding nucleotide" <221> Modified base <222> 20 <223> / Modified base = "3'-3'-linked deoxythymidine" <400> 347 gaggcuguag gcauaaauat 20 <210> 348 <211> 21 <212> DNA <213> Artificial sequence <223> Description of artificial sequence: sense strand of dsRNA <221> Modified base <222> 1 <223> / Modified base = "nucleotide: lacking 5'-phosphate group" <221> Modified base <222> 1, 2, 3, 4, 6, 9, 11, 12, 13, 14, 16, 17, 19 <223> / Modified base = "2'-O-methyl corresponding nucleotide" <221> Modified base <222> 21 <223> / Modified base = "5'-thio-phosphate thymidine" <221> Modified base <222> 5, 7, 8, 10, 15, 18 <223> / Modified base = "2'-hydroxy corresponding nucleotide" <400> 348 gucugcggcg uuuuaucaut t 21 <210> 349 <211> 21 <212> DNA <213> Artificial sequence <223> Description of artificial sequence: sense strand of dsRNA <221> Modified base <222> 1 <223> / Modified base = "nucleotide: lacking 5'-phosphate group" <221> Modified base <222> 1, 2, 4, 5, 6, 7, 8, 9, 10, 12, 13, 14, 15, 16, 18, 19 <223> / Modified base = "2'-O-methyl corresponding nucleotide" <221> Modified base <222> 21 <223> / Modified base = "5'-thio-phosphate thymidine" <221> Modified base <222> 3, 11, 17 <223> / Modified base = "2'-hydroxy corresponding nucleotide" <400> 349 caacuuuuuc accucugcct t 21 <210> 350 <211> 21 <212> DNA <213> Artificial sequence <223> Artificial sequence description: sense strand of dsRNA <221> Modified base <222> 1 <223> / Modified base = "Nucleotide: lacking 5'-phosphate group" <221> Modified base <222> 1, 2, 4, 6, 8, 10, 11, 12, 13, 15, 16, 17, 18 <223> / Modified base = "2'-O-methyl corresponding nucleotide" <221> Modified base <222> 21 <223> / Modified base = "5'-thiophosphate thymidine" <221> Modified base <222> 3, 5, 7, 9, 14, 19 <223> / Modified base = "2'-hydroxy corresponding nucleotide" <400> 350 ccgugugcac uucgcuucat t 21 <210> 351 <211> 20 <212> DNA <213> Artificial sequence <223> Artificial sequence description: sense strand of dsRNA <221> Modified base <222> 1 <223> / Modified base = "Nucleotide: lacking 5'-phosphate group" <221> Modified base <222> 2, 4, 6, 8, 10, 12, 14, 16, 18 <223> / Modified base = "2'-O-methyl corresponding nucleotide" <221> Modified base <222> 1, 3, 5, 7, 9, 11, 13, 15, 17, 19 <223> / Modified base = "2'-deoxy-2'-fluoro corresponding nucleotide" <221> Modified base <222> 20 <223> / Modified base = "3'-3'-linked deoxythymidine" <400> 351 ccgugugcac uucgcuucat 20 <210> 352 <211> 20 <212> DNA <213> Artificial sequence <223> Artificial sequence description: Sense strand of dsRNA <221> Modified base <222> 1 <223> / Modified base = "Nucleotide: lacking 5'-phosphate group" <221> Modified base <222> 2, 4, 6, 8, 10, 12, 14, 16, 18 <223> / Modified base = "2'-O-methyl corresponding nucleotide" <221> Modified base <222> 1, 3, 5, 7, 9, 11, 13, 15, 17, 19 <223> / Modified base = "2'-deoxy-2'-fluoro corresponding nucleotide" <221> Modified base <222> 20 <223> / Modified base = "3'-3'-linked deoxythymidine" <400> 352 ucaagguaug uugcccguat 20 <210> 353 <211> 20 <212> DNA <213> Artificial sequence <223> Artificial sequence description: Sense strand of dsRNA <221> Modified base <222> 1 <223> / Modified base = "Nucleotide: lacking 5'-phosphate group" <221> Modified base <222> 2, 4, 6, 8, 10, 12, 14, 16, 18 <223> / Modified base = "2'-O-methyl corresponding nucleotide" <221> Modified base <222> 1, 3, 5, 7, 9, 11, 13, 15, 17, 19 <223> / Modified base = "2'-deoxy-2'-fluoro corresponding nucleotide" <221> Modified base <222> 20 <223> / Modified base = "3'-3'-linked deoxythymidine" <400> 353 caguuuacua gugccauuat 20 <210> 354 <211> 21 <212> DNA <213> Artificial sequence <223> Description of artificial sequence: sense strand of dsRNA <221> Modified base <222> 1 <223> / Modified base = "Nucleotide: lacking 5'-phosphate group" <221> Modified base <222> 1, 2, 4, 8, 9, 10, 11, 12, 13, 14, 15, 18, 19 <223> / Modified base = "2'-O-methyl corresponding nucleotide" <221> Modified base <222> 21 <223> / Modified base = "5'-thio-phosphate thymidine" <221> Modified base <222> 3, 5, 6, 7, 16, 17 <223> / Modified base = "2'-hydroxy corresponding nucleotide" <400> 354 ugguggacuu cucucaauut t 21 <210> 355 <211> 20 <212> DNA <213> Artificial sequence <223> Description of artificial sequence: Sense strand of dsRNA <221> Modified base <222> 1 <223> / Modified base = "Nucleotide: lacking 5'-phosphate group" <221> Modified base <222> 2, 4, 6, 8, 10, 12, 14, 16, 18 <223> / Modified base = "2'-O-methyl corresponding nucleotide" <221> Modified base <222> 1, 3, 5, 7, 9, 11, 13, 15, 17, 19 <223> / Modified base = "2'-deoxy-2'-fluoro corresponding nucleotide" <221> Modified base <222> 20 <223> / Modified base = "3'-3'-linked deoxythymidine" <400> 355 agguauguug cccguuugat 20 <210> 356 <211> 21 <212> DNA <213> Artificial sequence <223> Description of artificial sequence: Sense strand of dsRNA <221> Modified base <222> 1 <223> / Modified base = "Nucleotide: lacking 5'-phosphate group" <221> Modified base <222> 1, 2, 4, 5, 6, 8, 10, 11, 13, 17 <223> / Modified base = "2'-O-methyl corresponding nucleotide" <221> Modified base <222> 21 <223> / Modified base = "5'-thiolated thymidine" <221> Modified base <222> 3, 7, 9, 12, 14, 15, 16, 18, 19 <223> / Modified base = "2'-hydroxy corresponding nucleotide" <400> 356 cugcucgugu uacaggcggt t 21 <210> 357 <211> 21 <212> DNA <213> Artificial sequence <223> Artificial sequence description: Sense strand of dsRNA <221> Modified base <222> 1 <223> / Modified base = "Nucleotide: lacking 5'-phosphate group" <221> Modified base <222> 1, 2, 3, 5, 6, 8, 9, 10, 12, 13, 14, 16, 17, 18, 19 <223> / Modified base = "2'-O-methyl corresponding nucleotide" <221> Modified base <222> 21 <223> / Modified base = "5'-thiolated thymidine" <221> Modified base <222> 4, 7, 11, 15 <223> / Modified base = "2'-hydroxy corresponding nucleotide" <400> 357 uauguugccc guuuguccut t 21 <210> 358 <211> 21 <212> DNA <213> Artificial sequence <223> Artificial sequence description: Sense strand of dsRNA <221> Modified base <222> 1 <223> / Modified base = "Nucleotide: lacking 5'-phosphate group" <221> Modified base <222> 1, 2, 7, 9, 11, 12, 14, 15, 16, 18, 19 <223> / Modified base = "2'-O-methyl corresponding nucleotide" <221> Modified base <222> 21 <223> / Modified base = "5'-thio-phosphate thymidine" <221> Modified base <222> 3, 4, 5, 6, 8, 10, 13, 17 <223> / Modified base = "2'-hydroxy corresponding nucleotide" <400> 358 ucaagguaug uugcccguut t 21 <210> 359 <211> 21 <212> DNA <213> Artificial sequence <223> Artificial sequence description: Sense strand of dsRNA <221> Modified base <222> 1 <223> / Modified base = "Nucleotide: lacking 5'-phosphate group" <221> Modified base <222> 1, 2, 3, 4, 6, 7, 10, 12, 13, 14, 15, 16 <223> / Modified base = "2'-O-methyl corresponding nucleotide" <221> Modified base <222> 21 <223> / Modified base = "5'-thio-phosphate thymidine" <221> Modified base <222> 5, 8, 9, 11, 17, 18, 19 <223> / Modified base = "2'-hydroxy corresponding nucleotide" <400> 359 ucuuaucaac acuuccggat t 21 <210> 360 <211> 20 <212> DNA <213> Artificial sequence <223> Description of artificial sequence: sense strand of dsRNA <221> Modified base <222> 1 <223> / Modified base = "Nucleotide: lacking 5'-phosphate group" <221> Modified base <222> 2, 4, 6, 8, 10, 12, 14, 16, 18 <223> / Modified base = "2'-O-methyl corresponding nucleotide" <221> Modified base <222> 1, 3, 5, 7, 9, 11, 13, 15, 17, 19 <223> / Modified base = "2'-deoxy-2'-fluoro corresponding nucleotide" <221> Modified base <222> 20 <223> / Modified base = "3'-3'-linked deoxythymidine" <400> 360 ucuuaucaac acuuccggat 20 <210> 361 <211> 21 <212> DNA <213> Artificial sequence <223> Description of artificial sequence: sense strand of dsRNA <221> Modified base <222> 1 <223> / Modified base = "Nucleotide: lacking 5'-phosphate group" <221> Modified base <222> 1, 2, 3, 4, 5, 6, 7, 9, 10, 11, 14, 15, 17, 18, 19 <223> / Modified base = "2'-O-methyl corresponding nucleotide" <221> Modified base <222> 21 <223> / Modified base = "5'-thiophosphate thymidine" <221> Modified base <222> 8, 12, 13, 16 <223> / Modified base = "2'-hydroxy corresponding nucleotide" <400> 361 caccucugcc uaaucaucut t 21 <210> 362 <211> 21 <212> DNA <213> Artificial sequence <223> Artificial sequence description: Sense strand of dsRNA <221> Modified base <222> 1 <223> / Modified base = "Nucleotide: lacking 5'-phosphate group" <221> Modified base <222> 1, 2, 10, 11, 12, 13, 14, 18, 19 <223> / Modified base = "2'-O-methyl corresponding nucleotide" <221> Modified base <222> 21 <223> / Modified base = "5'-thiophosphate thymidine" <221> Modified base <222> 3, 4, 5, 6, 7, 8, 9, 15, 16, 17 <223> / Modified base = "2'-hydroxy corresponding nucleotide" <400> 362 auaagaggac ucuuggacut t 21 <210> 363 <211> 20 <212> DNA <213> Artificial sequence <223> Artificial sequence description: Sense strand of dsRNA <221> Modified base <222> 1 <223> / Modified base = "Nucleotide: lacking 5'-phosphate group" <221> Modified base <222> 2, 4, 6, 8, 10, 12, 14, 16, 18 <223> / Modified base = "2'-O-methyl corresponding nucleotide" <221> Modified base <222> 1, 3, 5, 7, 9, 11, 13, 15, 17, 19 <223> / Modified base = "2'-deoxy-2'-fluoro corresponding nucleotide" <221> Modified base <222> 20 <223> / Modified base = "3'-3'-linked deoxythymidine" <400> 363 gucugcggcg uuuuaucaat 20 <210> 364 <211> 21 <212> DNA <213> Artificial sequence <223> Artificial sequence description: Sense strand of dsRNA <221> Modified base <222> 1 <223> / Modified base = "Nucleotide: lacking 5'-phosphate group" <221> Modified base <222> 1, 2, 3, 5, 6, 10, 11, 12, 13, 15, 19 <223> / Modified base = "2'-O-methyl corresponding nucleotide" <221> Modified base <222> 21 <223> / Modified base = "5'-thiophosphate thymidine" <221> Modified base <222> 4, 7, 8, 9, 14, 16, 17, 18 <223> / Modified base = "2'-hydroxy corresponding nucleotide" <400> 364 ggcgcugaau cccgcggact t 21 <210> 365 <211> 21 <212> DNA <213> Artificial sequence <223> Artificial sequence description: Sense strand of dsRNA <221> Modified base <222> 1 <223> / Modified base = "Nucleotide: lacking 5'-phosphate group" <221> Modified base <222> 1, 2, 3, 5, 6, 8, 15, 16, 17, 18 <223> / Modified base = "2'-O-methyl corresponding nucleotide" <221> Modified base <222> 21 <223> / Modified base = "5'-thio-phosphate thymidine" <221> Modified base <222> 4, 7, 9, 10, 11, 12, 13, 14, 19 <223> / Modified base = "2'-hydroxy corresponding nucleotide" <400> 365 cgcgucgcag aagaucucat t 21 <210> 366 <211> 21 <212> DNA <213> Artificial sequence <223> Artificial sequence description: sense strand of dsRNA <221> Modified base <222> 1 <223> / Modified base = "Nucleotide: lacking 5'-phosphate group" <221> Modified base <222> 1, 2, 3, 5, 6, 9, 12, 13, 16, 17, 18, 19 <223> / Modified base = "2'-O-methyl corresponding nucleotide" <221> Modified base <222> 21 <223> / Modified base = "5'-thio-phosphate thymidine" <221> Modified base <222> 4, 7, 8, 10, 11, 14, 15 <223> / Modified base = "2'-hydroxy corresponding nucleotide" <400> 366 aaugucaacg accgaccuut t 21 <210> 367 <211> 21 <212> DNA <213> Artificial sequence <223> Description of artificial sequence: sense strand of dsRNA <221> Modified base <222> 1 <223> / Modified base = "nucleotide: lacking 5'-phosphate group" <221> Modified base <222> 1, 2, 3, 4, 7, 8, 9, 11, 12, 15, 17, 18 <223> / Modified base = "2'-O-methyl corresponding nucleotide" <221> Modified base <222> 21 <223> / Modified base = "5'-thiophosphate thymidine" <221> Modified base <222> 5, 6, 10, 13, 14, 16, 19 <223> / Modified base = "2'-hydroxy corresponding nucleotide" <400> 367 gcucaguuua cuagugccat t 21 <210> 368 <211> 20 <212> DNA <213> Artificial sequence <223> Description of artificial sequence: sense strand of dsRNA <221> Modified base <222> 1 <223> / Modified base = "nucleotide: lacking 5'-phosphate group" <221> Modified base <222> 2, 4, 6, 8, 10, 12, 14, 16, 18 <223> / Modified base = "2'-O-methyl corresponding nucleotide" <221> Modified base <222> 1, 3, 5, 7, 9, 11, 13, 15, 17, 19 <223> / Modified base = "2'-deoxy-2'-fluoro corresponding nucleotide" <221> Modified base <222> 20 <223> / Modified base = "3'-3'-linked deoxythymidine" <400> 368 ugguggacuu cucucaauat 20 <210> 369 <211> 21 <212> DNA <213> Artificial sequence <223> Artificial sequence description: Sense strand of dsRNA <221> Modified base <222> 1 <223> / Modified base = "Nucleotide: lacking 5'-phosphate group" <221> Modified base <222> 1, 2, 3, 5, 6, 8, 10, 11, 13 <223> / Modified base = "2'-O-methyl corresponding nucleotide" <221> Modified base <222> 21 <223> / Modified base = "5'-thiophosphate thymidine" <221> Modified base <222> 4, 7, 9, 12, 14, 15, 16, 17, 18, 19 <223> / Modified base = "2'-hydroxy corresponding nucleotide" <400> 369 aucgccgcgu cgcagaagat t 21 <210> 370 <211> 21 <212> DNA <213> Artificial sequence <223> Artificial sequence description: Sense strand of dsRNA <221> Modified base <222> 1 <223> / Modified base = "Nucleotide: lacking 5'-phosphate group" <221> Modified base <222> 1, 2, 3, 5, 6, 7, 9, 10, 11, 14, 17, 18, 19 <223> / Modified base = "2'-O-methyl corresponding nucleotide" <221> Modified base <222> 21 <223> / Modified base = "5'-thio-phosphate thymidine" <221> Modified base <222> 4, 8, 12, 13, 15, 16 <223> / Modified base = "2'-hydroxy corresponding nucleotide" <400> 370 gccauuuguu cagugguuct t 21 <210> 371 <211> 21 <212> DNA <213> Artificial sequence <223> Artificial sequence description: sense strand of dsRNA <221> Modified base <222> 1 <223> / Modified base = "Nucleotide: lacking 5'-phosphate group" <221> Modified base <222> 1, 2, 4, 5, 6, 8, 10, 11, 13, 18, 19 <223> / Modified base = "2'-O-methyl corresponding nucleotide" <221> Modified base <222> 21 <223> / Modified base = "5'-thio-phosphate thymidine" <221> Modified base <222> 3, 7, 9, 12, 14, 15, 16, 17 <223> / Modified base = "2'-hydroxy corresponding nucleotide" <400> 371 cgauccauac ugcggaacut t 21 <210> 372 <211> 21 <212> DNA <213> Artificial sequence <223> Description of artificial sequence: sense strand of dsRNA <221> Modified base <222> 1 <223> / Modified base = "nucleotide: lacking 5'-phosphate group" <221> Modified base <222> 1, 2, 4, 5, 6, 7, 8, 10, 11, 12, 15, 16, 18, 19 <223> / Modified base = "2'-O-methyl corresponding nucleotide" <221> Modified base <222> 21 <223> / Modified base = "5'-thio-phosphate thymidine" <221> Modified base <222> 3, 9, 13, 14, 17 <223> / Modified base = "2'-hydroxy corresponding nucleotide" <400> 372 ucaccucugc cuaaucauct t 21 <210> 373 <211> 21 <212> DNA <213> Artificial sequence <223> Description of artificial sequence: sense strand of dsRNA <221> Modified base <222> 1 <223> / Modified base = "nucleotide: lacking 5'-phosphate group" <221> Modified base <222> 1, 2, 6, 7, 8, 9, 10, 11, 12, 13, 16, 17, 18, 19 <223> / Modified base = "2'-O-methyl corresponding nucleotide" <221> Modified base <222> 21 <223> / Modified base = "5'-thio-phosphate thymidine" <221> Modified base <222> 3, 4, 5, 14, 15 <223> / Modified base = "2'-hydroxy corresponding nucleotide" <400> 373 guggacuucu cucaauuuut t 21 <210> 374 <211> 21 <212> DNA <213> Artificial sequence <223> Artificial sequence description: Sense strand of dsRNA <221> Modified base <222> 1 <223> / Modified base = "Nucleotide: lacking 5'-phosphate group" <221> Modified base <222> 1, 2, 4, 5, 7, 8, 10, 12, 13, 14, 15, 16 <223> / Modified base = "2'-O-methyl corresponding nucleotide" <221> Modified base <222> 21 <223> / Modified base = "5'-thiophosphate thymidine" <221> Modified base <222> 3, 6, 9, 11, 17, 18, 19 <223> / Modified base = "2'-hydroxy corresponding nucleotide" <400> 374 gggucaccau auucuugggt t 21 <210> 375 <211> 21 <212> DNA <213> Artificial sequence <223> Artificial sequence description: Sense strand of dsRNA <221> Modified base <222> 1 <223> / Modified base = "Nucleotide: lacking 5'-phosphate group" <221> Modified base <222> 1, 2, 3, 5, 7, 8, 10, 17, 18, 19 <223> / Modified base = "2'-O-methyl corresponding nucleotide" <221> Modified base <222> 21 <223> / Modified base = "5'-thio-phosphate thymidine" <221> Modified base <222> 4, 6, 9, 11, 12, 13, 14, 15, 16 <223> / Modified base = "2'-hydroxy corresponding nucleotide" <400> 375 gccgcgucgc agaagaucut t 21 <210> 376 <211> 21 <212> DNA <213> Artificial sequence <223> Artificial sequence description: Sense strand of dsRNA <221> Modified base <222> 1 <223> / Modified base = "Nucleotide: lacking 5'-phosphate group" <221> Modified base <222> 1, 2, 5, 6, 8, 9, 11, 13, 14, 16 <223> / Modified base = "2'-O-methyl corresponding nucleotide" <221> Modified base <222> 21 <223> / Modified base = "5'-thio-phosphate thymidine" <221> Modified base <222> 3, 4, 7, 10, 12, 15, 17, 18, 19 <223> / Modified base = "2'-hydroxy corresponding nucleotide" <400> 376 ucaaucgccg cgucgcagat t 21 <210> 377 <211> 21 <212> DNA <213> Artificial Sequence <223> Description of artificial sequence: sense strand of dsRNA <221> Modified base <222> 1 <223> / Modified base = "nucleotide: lacking 5'-phosphate group" <221> Modified base <222> 1, 2, 5, 7, 9, 10, 11, 13, 16, 18, 19 <223> / Modified base = "2'-O-methyl corresponding nucleotide" <221> Modified base <222> 21 <223> / Modified base = "5'-thiophosphate thymidine" <221> Modified base <222> 3, 4, 6, 8, 12, 14, 15, 17 <223> / Modified base = "2'-hydroxy corresponding nucleotide" <400> 377 uggauguguc ugcggcguut t 21 <210> 378 <211> 21 <212> DNA <213> Artificial Sequence <223> Description of artificial sequence: sense strand of dsRNA <221> Modified base <222> 1 <223> / Modified base = "nucleotide: lacking 5'-phosphate group" <221> Modified base <222> 1, 2, 3, 4, 6, 7, 8, 12, 13, 14, 15, 16 <223> / Modified base = "2'-O-methyl corresponding nucleotide" <221> Modified base <222> 21 <223> / Modified base = "5'-thiophosphate thymidine" <221> Modified base <222> 5, 9, 10, 11, 17, 18, 19 <223> / Modified base = "2'-hydroxy corresponding nucleotide" <400> 378 uacuguucaa gccuccaagt t 21 <210> 379 <211> 21 <212> DNA <213> Artificial sequence <223> Artificial sequence description: Sense strand of dsRNA <221> Modified base <222> 1 <223> / Modified base = "Nucleotide: lacking 5'-phosphate group" <221> Modified base <222> 1, 2, 3, 4, 6, 7, 10, 12, 13, 15, 16, 17, 19 <223> / Modified base = "2'-O-methyl corresponding nucleotide" <221> Modified base <222> 21 <223> / Modified base = "5'-thiophosphate thymidine" <221> Modified base <222> 5, 8, 9, 11, 14, 18 <223> / Modified base = "2'-hydroxy corresponding nucleotide" <400> 379 guuuacuagu gccauuugut t 21 <210> 380 <211> 21 <212> DNA <213> Artificial sequence <223> Artificial sequence description: Sense strand of dsRNA <221> Modified base <222> 1 <223> / Modified base = "Nucleotide: lacking 5'-phosphate group" <221> Modified base <222> 1, 2, 3, 6, 8, 9, 11, 12, 13, 15, 16, 17 <223> / Modified base = "2'-O-methyl corresponding nucleotide" <221> Modified base <222> 21 <223> / Modified base = "5'-thio-phosphate thymidine" <221> Modified base <222> 4, 5, 7, 10, 14, 18, 19 <223> / Modified base = "2'-hydroxy corresponding nucleotide" <400> 380 acuagugcca uuuguucagt t 21 <210> 381 <211> 21 <212> DNA <213> Artificial sequence <223> Artificial sequence description: Sense strand of dsRNA <221> Modified base <222> 1 <223> / Modified base = "Nucleotide: lacking 5'-phosphate group" <221> Modified base <222> 1, 2, 4, 6, 7, 9, 16, 17, 18, 19 <223> / Modified base = "2'-O-methyl corresponding nucleotide" <221> Modified base <222> 21 <223> / Modified base = "5'-thio-phosphate thymidine" <221> Modified base <222> 3, 5, 8, 10, 11, 12, 13, 14, 15 <223> / Modified base = "2'-hydroxy corresponding nucleotide" <400> 381 ccgcgucgca gaagaucuct t 21 <210> 382 <211> 21 <212> DNA <213> Artificial sequence <223> Artificial sequence description: Sense strand of dsRNA <221> Modified base <222> 1 <223> / Modified base = "Nucleotide: lacking 5'-phosphate group" <221> Modified base <222> 1, 2, 3, 4, 5, 6, 8, 9, 12, 14, 15, 16, 17, 18 <223> / Modified base = "2'-O-methyl corresponding nucleotide" <221> Modified base <222> 21 <223> / Modified base = "5'-thio-phosphate thymidine" <221> Modified base <222> 7, 10, 11, 13, 19 <223> / Modified base = "2'-hydroxy corresponding nucleotide" <400> 382 uaucuuauca acacuuccgt t 21 <210> 383 <211> 21 <212> DNA <213> Artificial sequence <223> Artificial sequence description: sense strand of dsRNA <221> Modified base <222> 1 <223> / Modified base = "Nucleotide: lacking 5'-phosphate group" <221> Modified base <222> 1, 2, 3, 4, 9, 10, 11, 13, 16, 17, 18, 19 <223> / Modified base = "2'-O-methyl corresponding nucleotide" <221> Modified base <222> 21 <223> / Modified base = "5'-thio-phosphate thymidine" <221> Modified base <222> 5, 6, 7, 8, 12, 14, 15 <223> / Modified base = "2'-hydroxy corresponding nucleotide" <400> 383 ggccaaaauu cgcaguccct t 21 <210> 384 <211> 21 <212> DNA <213> Artificial sequence <223> Description of artificial sequence: sense strand of dsRNA <221> Modified base <222> 1 <223> / Modified base = "Nucleotide: lacking 5'-phosphate group" <221> Modified base <222> 1, 2, 3, 5, 6, 7, 8, 9, 11, 12, 13, 16, 17, 19 <223> / Modified base = "2'-O-methyl corresponding nucleotide" <221> Modified base <222> 21 <223> / Modified base = "5'-thio-phosphate thymidine" <221> Modified base <222> 4, 10, 14, 15, 18 <223> / Modified base = "2'-hydroxy corresponding nucleotide" <400> 384 uucaccucug ccuaaucaut t 21 <210> 385 <211> 21 <212> DNA <213> Artificial sequence <223> Description of artificial sequence: sense strand of dsRNA <221> Modified base <222> 1 <223> / Modified base = "Nucleotide: lacking 5'-phosphate group" <221> Modified base <222> 1, 2, 3, 6, 7, 8, 10, 11, 14, 16, 17, 19 <223> / Modified base = "2'-O-methyl corresponding nucleotide" <221> Modified base <222> 21 <223> / Modified base = "5'-thio-phosphate thymidine" <221> Modified base <222> 4, 5, 9, 12, 13, 15, 18 <223> / Modified base = "2'-hydroxy corresponding nucleotide" <400> 385 cucaguuuac uagugccaut t 21 <210> 386 <211> 21 <212> DNA <213> Artificial sequence <223> Artificial sequence description: Sense strand of dsRNA <221> Modified base <222> 1 <223> / Modified base = "Nucleotide: lacking 5'-phosphate group" <221> Modified base <222> 1, 2, 3, 4, 6, 7, 8, 10, 11, 12, 14, 15, 16, 17, 18, 19 <223> / Modified base = "2'-O-methyl corresponding nucleotide" <221> Modified base <222> 21 <223> / Modified base = "5'-thio-phosphate thymidine" <221> Modified base <222> 5, 9, 13 <223> / Modified base = "2'-hydroxy corresponding nucleotide" <400> 386 uguugcccgu uuguccucut t 21 <210> 387 <211> 21 <212> DNA <213> Artificial sequence <223> Artificial sequence description: Sense strand of dsRNA <221> Modified base <222> 1 <223> / Modified base = "Nucleotide: lacking 5'-phosphate group" <221> Modified base <222> 1, 2, 4, 6, 7, 9, 10, 11, 13, 14, 15, 18 <223> / Modified base = "2'-O-methyl corresponding nucleotide" <221> Modified base <222> 21 <223> / Modified base = "5'-thiophosphate thymidine" <221> Modified base <222> 3, 5, 8, 12, 16, 17, 19 <223> / Modified base = "2'-hydroxy corresponding nucleotide" <400> 387 uagugccauu uguucagugt t 21 <210> 388 <211> 21 <212> DNA <213> Artificial sequence <223> Description of artificial sequence: Sense strand of dsRNA <221> Modified base <222> 1 <223> / Modified base = "Nucleotide: lacking 5'-phosphate group" <221> Modified base <222> 1, 2, 4, 5, 7, 11, 13, 17, 18 <223> / Modified base = "2'-O-methyl corresponding nucleotide" <221> Modified base <222> 21 <223> / Modified base = "5'-thiophosphate thymidine" <221> Modified base <222> 3, 6, 8, 9, 10, 12, 14, 15, 16, 19 <223> / Modified base = "2'-hydroxy corresponding nucleotide" <400> 388 aggcuguagg cauaaauugt t 21 <210> 389 <211> 21 <212> DNA <213> Artificial sequence <223> Description of artificial sequence: sense strand of dsRNA <221> Modified base <222> 1 <223> / Modified base = "nucleotide: lacking 5'-phosphate group" <221> Modified base <222> 1, 2, 4, 6, 7, 8, 10, 13, 15, 16, 17, 18 <223> / Modified base = "2'-O-methyl corresponding nucleotide" <221> Modified base <222> 21 <223> / Modified base = "5'-thio-phosphate thymidine" <221> Modified base <222> 3, 5, 9, 11, 12, 14, 19 <223> / Modified base = "2'-hydroxy corresponding nucleotide" <400> 389 augugucugc ggcguuuuat t 21 <210> 390 <211> 20 <212> DNA <213> Artificial sequence <223> Description of artificial sequence: sense strand of dsRNA <221> Modified base <222> 1 <223> / Modified base = "nucleotide: lacking 5'-phosphate group" <221> Modified base <222> 2, 4, 6, 8, 10, 12, 14, 16, 18 <223> / Modified base = "2'-O-methyl corresponding nucleotide" <221> Modified base <222> 1, 3, 5, 7, 9, 11, 13, 15, 17, 19 <223> / Modified base = "2'-deoxy-2'-fluoro corresponding nucleotide" <221> Modified base <222> 20 <223> / Modified base = "3'-3'-linked deoxythymidine" <400> 390 augugucugc ggcguuuuat 20 <210> 391 <211> 21 <212> DNA <213> Artificial sequence <223> Artificial sequence description: Sense strand of dsRNA <221> Modified base <222> 1 <223> / Modified base = "Nucleotide: lacking 5'-phosphate group" <221> Modified base <222> 1, 2, 3, 4, 5, 7, 8, 9, 10, 12, 13, 14, 15, 16, 18 <223> / Modified base = "2'-O-methyl corresponding nucleotide" <221> Modified base <222> 21 <223> / Modified base = "5'-thio-phosphate thymidine" <221> Modified base <222> 6, 11, 17, 19 <223> / Modified base = "2'-hydroxy corresponding nucleotide" <400> 391 acuucgcuuc accucugcat t 21 <210> 392 <211> 21 <212> DNA <213> Artificial sequence <223> Artificial sequence description: Sense strand of dsRNA <221> Modified base <222> 1 <223> / Modified base = "Nucleotide: lacking 5'-phosphate group" <221> Modified base <222> 1, 2, 3, 5, 7, 9, 10, 11, 12, 14, 15, 16, 17, 19 <223> / Modified base = "2'-O-methyl corresponding nucleotide" <221> Modified base <222> 21 <223> / Modified base = "5'-thiophosphate thymidine" <221> Modified base <222> 4, 6, 8, 13, 18 <223> / Modified base = "2'-hydroxy corresponding nucleotide" <400> 392 cgugugcacu ucgcuucact t 21 <210> 393 <211> 21 <212> DNA <213> Artificial sequence <223> Artificial sequence description: Sense strand of dsRNA <221> Modified base <222> 1 <223> / Modified base = "Nucleotide: lacking 5'-phosphate group" <221> Modified base <222> 1, 2, 5, 9, 10, 11, 12, 13, 14, 15, 16, 19 <223> / Modified base = "2'-O-methyl corresponding nucleotide" <221> Modified base <222> 21 <223> / Modified base = "5'-thiophosphate thymidine" <221> Modified base <222> 3, 4, 6, 7, 8, 17, 18 <223> / Modified base = "2'-hydroxy corresponding nucleotide" <400> 393 gugguggacu ucucucaaut t 21 <210> 394 <211> 21 <212> DNA <213> Artificial sequence <223> Artificial sequence description: Sense strand of dsRNA <221> Modified base <222> 1 <223> / Modified base = "Nucleotide: lacking 5'-phosphate group" <221> Modified base <222> 1, 2, 3, 5, 6, 7, 9, 12, 14, 15, 16, 17, 19 <223> / Modified base = "2'-O-methyl corresponding nucleotide" <221> Modified base <222> 21 <223> / Modified base = "5'-thiophosphate thymidine" <221> Modified base <222> 4, 8, 10, 11, 13, 18 <223> / Modified base = "2'-hydroxy corresponding nucleotide" <400> 394 ugugucugcg gcguuuuaut t 21 <210> 395 <211> 21 <212> DNA <213> Artificial sequence <223> Artificial sequence description: sense strand of dsRNA <221> Modified base <222> 1 <223> / Modified base = "Nucleotide: lacking 5'-phosphate group" <221> Modified base <222> 1, 2, 5, 7, 9, 10, 12, 13, 14, 16, 17, 18 <223> / Modified base = "2'-O-methyl corresponding nucleotide" <221> Modified base <222> 21 <223> / Modified base = "5'-thiophosphate thymidine" <221> Modified base <222> 3, 4, 6, 8, 11, 15, 19 <223> / Modified base = "2'-hydroxy corresponding nucleotide" <400> 395 aagguauguu gcccguuugt t 21 <210> 396 <211> 21 <212> DNA <213> Artificial sequence <223> Artificial sequence description: sense strand of dsRNA <221> Modified base <222> 1 <223> / Modified base = "Nucleotide: lacking 5'-phosphate group" <221> Modified base <222> 1, 2, 5, 8, 9, 12, 13, 14, 15 <223> / Modified base = "2'-O-methyl corresponding nucleotide" <221> Modified base <222> 21 <223> / Modified base = "5'-thiophosphate thymidine" <221> Modified base <222> 3, 4, 6, 7, 10, 11, 16, 17, 18, 19 <223> / Modified base = "2'-hydroxy corresponding nucleotide" <400> 396 ucaacgaccg accuugaggt t 21 <210> 397 <211> 21 <212> DNA <213> Artificial sequence <223> Artificial sequence description: sense strand of dsRNA <221> Modified base <222> 1 <223> / Modified base = "Nucleotide: lacking 5'-phosphate group" <221> Modified base <222> 1, 2, 3, 11, 12, 13, 14, 15, 19 <223> / Modified base = "2'-O-methyl corresponding nucleotide" <221> Modified base <222> 21 <223> / Modified base = "5'-thiophosphate thymidine" <221> Modified base <222> 4, 5, 6, 7, 8, 9, 10, 16, 17, 18 <223> / Modified base = "2'-hydroxy corresponding nucleotide" <400> 397 cauaagagga cucuuggact t 21 <210> 398 <211> 21 <212> DNA <213> Artificial sequence <223> Artificial sequence description: Sense strand of dsRNA <221> Modified base <222> 1 <223> / Modified base = "Nucleotide: lacking 5'-phosphate group" <221> Modified base <222> 1, 2, 3, 6, 9, 10, 13, 14, 15, 16 <223> / Modified base = "2'-O-methyl corresponding nucleotide" <221> Modified base <222> 21 <223> / Modified base = "5'-thiophosphate thymidine" <221> Modified base <222> 4, 5, 7, 8, 11, 12, 17, 18, 19 <223> / Modified base = "2'-hydroxy corresponding nucleotide" <400> 398 gucaacgacc gaccuugagt t 21 <210> 399 <211> 21 <212> DNA <213> Artificial sequence <223> Artificial sequence description: Sense strand of dsRNA <221> Modified base <222> 1 <223> / Modified base = "Nucleotide: lacking 5'-phosphate group" <221> Modified base <222> 1, 2, 4, 5, 6, 7, 8, 14 <223> / Modified base = "2'-O-methyl corresponding nucleotide" <221> Modified base <222> 21 <223> / Modified base = "5'-thiophosphate thymidine" <221> Modified base <222> 3, 9, 10, 11, 12, 13, 15, 16, 17, 18, 19 <223> / Modified base = "2'-hydroxy corresponding nucleotide" <400> 399 auauucuugg gaacaagagt t 21 <210> 400 <211> 21 <212> DNA <213> Artificial sequence <223> Artificial sequence description: Sense strand of dsRNA <221> Modified base <222> 1 <223> / Modified base = "Nucleotide: lacking 5'-phosphate group" <221> Modified base <222> 1, 2, 3, 4, 5, 7, 9, 10, 12, 16 <223> / Modified base = "2'-O-methyl corresponding nucleotide" <221> Modified base <222> 21 <223> / Modified base = "5'-thiophosphate thymidine" <221> Modified base <222> 6, 8, 11, 13, 14, 15, 17, 18, 19 <223> / Modified base = "2'-hydroxy corresponding nucleotide" <400> 400 ugcucguguu acaggcgggt t 21 <210> 401 <211> 21 <212> DNA <213> Artificial Sequence <223> Description of artificial sequence: sense strand of dsRNA <221> Modified base <222> 1 <223> / Modified base = "Nucleotide: lacking 5'-phosphate group" <221> Modified base <222> 1, 2, 4, 5, 7, 8, 10, 12, 13, 15 <223> / Modified base = "2'-O-methyl corresponding nucleotide" <221> Modified base <222> 21 <223> / Modified base = "5'-thio-phosphate thymidine" <221> Modified base <222> 3, 6, 9, 11, 14, 16, 17, 18, 19 <223> / Modified base = "2'-hydroxy corresponding nucleotide" <400> 401 caaucgccgc gucgcagaat t 21 <210> 402 <211> 21 <212> DNA <213> Artificial Sequence <223> Description of artificial sequence: sense strand of dsRNA <221> Modified base <222> 1 <223> / Modified base = "Nucleotide: lacking 5'-phosphate group" <221> Modified base <222> 1, 2, 3, 5, 6, 7, 11, 12, 13, 14, 15, 19 <223> / Modified base = "2'-O-methyl corresponding nucleotide" <221> Modified base <222> 21 <223> / Modified base = "5'-thio-phosphate thymidine" <221> Modified base <222> 4, 8, 9, 10, 16, 17, 18 <223> / Modified base = "2'-hydroxy corresponding nucleotide" <400> 402 acuguucaag ccuccaagct t 21 <210> 403 <211> 21 <212> DNA <213> Artificial sequence <223> Artificial sequence description: Sense strand of dsRNA <221> Modified base <222> 1 <223> / Modified base = "Nucleotide: lacking 5'-phosphate group" <221> Modified base <222> 1, 2, 3, 4, 6, 8, 9, 11, 18, 19 <223> / Modified base = "2'-O-methyl corresponding nucleotide" <221> Modified base <222> 21 <223> / Modified base = "5'-thiophosphate thymidine" <221> Modified base <222> 5, 7, 10, 12, 13, 14, 15, 16, 17 <223> / Modified base = "2'-hydroxy corresponding nucleotide" <400> 403 cgccgcgucg cagaagauct t 21 <210> 404 <211> 21 <212> DNA <213> Artificial sequence <223> Artificial sequence description: Sense strand of dsRNA <221> Modified base <222> 1 <223> / Modified base = "Nucleotide: lacking 5'-phosphate group" <221> Modified base <222> 1, 2, 3, 4, 5, 7, 8, 9, 12, 15, 16, 17, 19 <223> / Modified base = "2'-O-methyl corresponding nucleotide" <221> Modified base <222> 21 <223> / Modified base = "5'-thio-phosphate thymidine" <221> Modified base <222> 6, 10, 11, 13, 14, 18 <223> / Modified base = "2'-hydroxy corresponding nucleotide" <400> 404 cauuuguuca gugguucgut t 21 <210> 405 <211> 21 <212> DNA <213> Artificial sequence <223> Artificial sequence description: Sense strand of dsRNA <221> Modified base <222> 1 <223> / Modified base = "Nucleotide: lacking 5'-phosphate group" <221> Modified base <222> 1, 2, 3, 4, 8, 9, 10, 11, 13, 17 <223> / Modified base = "2'-O-methyl corresponding nucleotide" <221> Modified base <222> 21 <223> / Modified base = "5'-thio-phosphate thymidine" <221> Modified base <222> 5, 6, 7, 12, 14, 15, 16, 18, 19 <223> / Modified base = "2'-hydroxy corresponding nucleotide" <400> 405 cgcugaaucc cgcggacgat t 21 <210> 406 <211> 21 <212> DNA <213> Artificial Sequence <223> Description of artificial sequence: sense strand of dsRNA <221> Modified base <222> 1 <223> / Modified base = "nucleotide: lacking 5'-phosphate group" <221> Modified base <222> 1, 2, 5, 6, 8, 9, 11, 13, 14, 15, 16, 17 <223> / Modified base = "2'-O-methyl corresponding nucleotide" <221> Modified base <222> 21 <223> / Modified base = "5'-thiophosphate thymidine" <221> Modified base <222> 3, 4, 7, 10, 12, 18, 19 <223> / Modified base = "2'-hydroxy corresponding nucleotide" <400> 406 ugggucacca uauucuuggt t 21 <210> 407 <211> 21 <212> DNA <213> Artificial Sequence <223> Description of artificial sequence: sense strand of dsRNA <221> Modified base <222> 1 <223> / Modified base = "nucleotide: lacking 5'-phosphate group" <221> Modified base <222> 1, 2, 3, 4, 5, 6, 8, 9, 12, 13, 14, 16, 18, 19 <223> / Modified base = "2'-O-methyl corresponding nucleotide" <221> Modified base <222> 21 <223> / Modified base = "5'-thiophosphate thymidine" <221> Modified base <222> 7, 10, 11, 15, 17 <223> / Modified base = "2'-hydroxy corresponding nucleotide" <400> 407 uccucugccg auccauacut t 21 <210> 408 <211> 21 <212> DNA <213> Artificial sequence <223> Artificial sequence description: sense strand of dsRNA <221> Modified base <222> 1 <223> / Modified base = "Nucleotide: lacking 5'-phosphate group" <221> Modified base <222> 1, 2, 4, 5, 8, 11, 12, 15, 16, 17, 18 <223> / Modified base = "2'-O-methyl corresponding nucleotide" <221> Modified base <222> 21 <223> / Modified base = "5'-thiophosphate thymidine" <221> Modified base <222> 3, 6, 7, 9, 10, 13, 14, 19 <223> / Modified base = "2'-hydroxy corresponding nucleotide" <400> 408 augucaacga ccgaccuugt t 21 <210> 409 <211> 21 <212> DNA <213> Artificial sequence <223> Artificial sequence description: sense strand of dsRNA <221> Modified base <222> 1 <223> / Modified base = "Nucleotide: lacking 5'-phosphate group" <221> Modified base <222> 1, 2, 3, 4, 5, 7, 8, 9, 12, 13, 15, 16, 17, 18 <223> / Modified base = "2'-O-methyl corresponding nucleotide" <221> Modified base <222> 21 <223> / Modified base = "5'-thiophosphate thymidine" <221> Modified base <222> 6, 10, 11, 14, 19 <223> / Modified base = "2'-hydroxy corresponding nucleotide" <400> 409 ccucugccua aucaucucat t 21 <210> 410 <211> 21 <212> DNA <213> Artificial sequence <223> Artificial sequence description: Sense strand of dsRNA <221> Modified base <222> 1 <223> / Modified base = "Nucleotide: lacking 5'-phosphate group" <221> Modified base <222> 1, 2, 3, 5, 7, 9, 11, 12, 13, 14, 16, 17, 18, 19 <223> / Modified base = "2'-O-methyl corresponding nucleotide" <221> Modified base <222> 21 <223> / Modified base = "5'-thiophosphate thymidine" <221> Modified base <222> 4, 6, 8, 10, 15 <223> / Modified base = "2'-hydroxy corresponding nucleotide" <400> 410 accgugugca cuucgcuuct t 21 <210> 411 <211> 21 <212> DNA <213> Artificial sequence <223> Artificial sequence description: Sense strand of dsRNA <221> Modified base <222> 1 <223> / Modified base = "Nucleotide: lacking 5'-phosphate group" <221> Modified base <222> 1, 2, 3, 4, 7, 8, 9, 11, 13, 14, 16 <223> / Modified base = "2'-O-methyl corresponding nucleotide" <221> Modified base <222> 21 <223> / Modified base = "5'-thio-phosphate thymidine" <221> Modified base <222> 5, 6, 10, 12, 15, 17, 18, 19 <223> / Modified base = "2'-hydroxy corresponding nucleotide" <400> 411 ugccgaucca uacugcggat t 21 <210> 412 <211> 21 <212> DNA <213> Artificial sequence <223> Artificial sequence description: Sense strand of dsRNA <221> Modified base <222> 1 <223> / Modified base = "Nucleotide: lacking 5'-phosphate group" <221> Modified base <222> 1, 2, 6, 7, 8, 12, 13, 14, 16, 19 <223> / Modified base = "2'-O-methyl corresponding nucleotide" <221> Modified base <222> 21 <223> / Modified base = "5'-thio-phosphate thymidine" <221> Modified base <222> 3, 4, 5, 9, 10, 11, 15, 17, 18 <223> / Modified base = "2'-hydroxy corresponding nucleotide" <400> 412 cagagucuag acucguggut t 21 <210> 413 <211> 21 <212> DNA <213> Artificial sequence <223> Description of artificial sequence: sense strand of dsRNA <221> Modified base <222> 1 <223> / Modified base = "nucleotide: lacking 5'-phosphate group" <221> Modified base <222> 1, 2, 4, 5, 6, 10, 11, 12, 13, 14, 18, 19 <223> / Modified base = "2'-O-methyl corresponding nucleotide" <221> Modified base <222> 21 <223> / Modified base = "5'-thio-phosphate thymidine" <221> Modified base <222> 3, 7, 8, 9, 15, 16, 17 <223> / Modified base = "2'-hydroxy corresponding nucleotide" <400> 413 cuguucaagc cuccaagcut t 21 <210> 414 <211> 21 <212> DNA <213> Artificial sequence <223> Description of artificial sequence: sense strand of dsRNA <221> Modified base <222> 1 <223> / Modified base = "nucleotide: lacking 5'-phosphate group" <221> Modified base <222> 1, 2, 6, 7, 9, 13, 15, 19 <223> / Modified base = "2'-O-methyl corresponding nucleotide" <221> Modified base <222> 21 <223> / Modified base = "5'-thiophosphate thymidine" <221> Modified base <222> 3, 4, 5, 8, 10, 11, 12, 14, 16, 17, 18 <223> / Modified base = "2'-hydroxy corresponding nucleotide" <400> 414 ggaggcugua ggcauaaaut t 21 <210> 415 <211> 21 <212> DNA <213> Artificial sequence <223> Artificial sequence description: Sense strand of dsRNA <221> Modified base <222> 1 <223> / Modified base = "Nucleotide: lacking 5'-phosphate group" <221> Modified base <222> 1, 2, 7, 8, 10, 14, 16 <223> / Modified base = "2'-O-methyl corresponding nucleotide" <221> Modified base <222> 21 <223> / Modified base = "5'-thiophosphate thymidine" <221> Modified base <222> 3, 4, 5, 6, 9, 11, 12, 13, 15, 17, 18, 19 <223> / Modified base = "2'-hydroxy corresponding nucleotide" <400> 415 aggaggcugu aggcauaaat t 21 <210> 416 <211> 21 <212> DNA <213> Artificial sequence <223> Artificial sequence description: Sense strand of dsRNA <221> Modified base <222> 1 <223> / Modified base = "Nucleotide: lacking 5'-phosphate group" <221> Modified base <222> 1, 2, 3, 7, 8, 9, 10, 11, 12, 13, 14, 17, 18, 19 <223> / Modified base = "2'-O-methyl corresponding nucleotide" <221> Modified base <222> 21 <223> / Modified base = "5'-thio-phosphate thymidine" <221> Modified base <222> 4, 5, 6, 15, 16 <223> / Modified base = "2'-hydroxy corresponding nucleotide" <400> 416 gguggacuuc ucucaauuut t 21 <210> 417 <211> 21 <212> DNA <213> Artificial sequence <223> Description of artificial sequence: Sense strand of dsRNA <221> Modified base <222> 1 <223> / Modified base = "Nucleotide: lacking 5'-phosphate group" <221> Modified base <222> 1, 2, 5, 6, 7, 8, 9, 10, 11, 13, 14, 15, 16, 17, 19 <223> / Modified base = "2'-O-methyl corresponding nucleotide" <221> Modified base <222> 21 <223> / Modified base = "5'-thio-phosphate thymidine" <221> Modified base <222> 3, 4, 12, 18 <223> / Modified base = "2'-hydroxy corresponding nucleotide" <400> 417 gcaacuuuuu caccucugct t 21 <210> 418 <211> 20 <212> DNA <213> Artificial sequence <223> Description of artificial sequence: sense strand of dsRNA <221> Modified base <222> 1 <223> / Modified base = "nucleotide: lacking 5'-phosphate group" <221> Modified base <222> 2, 4, 6, 8, 10, 12, 14, 16, 18 <223> / Modified base = "2'-O-methyl corresponding nucleotide" <221> Modified base <222> 1, 3, 5, 7, 9, 11, 13, 15, 17, 19 <223> / Modified base = "2'-deoxy-2'-fluoro corresponding nucleotide" <221> Modified base <222> 20 <223> / Modified base = "3'-3'-linked deoxythymidine" <400> 418 cugcucgugu uacaggcgat 20 <210> 419 <211> 21 <212> DNA <213> Artificial sequence <223> Description of artificial sequence: sense strand of dsRNA <221> Modified base <222> 1 <223> / Modified base = "nucleotide: lacking 5'-phosphate group" <221> Modified base <222> 1, 2, 5, 7, 8, 10, 11, 12, 14, 15, 16, 19 <223> / Modified base = "2'-O-methyl corresponding nucleotide" <221> Modified base <222> 21 <223> / Modified base = "5'-thio-phosphate thymidine" <221> Modified base <222> 3, 4, 6, 9, 13, 17, 18 <223> / Modified base = "2'-hydroxy corresponding nucleotide" <400> 419 cuagugccau uuguucagut t 21 <210> 420 <211> 21 <212> DNA <213> Artificial sequence <223> Artificial sequence description: Sense strand of dsRNA <221> Modified base <222> 1 <223> / Modified base = "Nucleotide: lacking 5'-phosphate group" <221> Modified base <222> 1, 2, 4, 5, 8, 9, 10, 12, 14, 15, 17 <223> / Modified base = "2'-O-methyl corresponding nucleotide" <221> Modified base <222> 21 <223> / Modified base = "5'-thiophosphate thymidine" <221> Modified base <222> 3, 6, 7, 11, 13, 16, 18, 19 <223> / Modified base = "2'-hydroxy corresponding nucleotide" <400> 420 cugccgaucc auacugcggt t 21 <210> 421 <211> 21 <212> DNA <213> Artificial sequence <223> Artificial sequence description: Sense strand of dsRNA <221> Modified base <222> 1 <223> / Modified base = "Nucleotide: lacking 5'-phosphate group" <221> Modified base <222> 1, 2, 4, 6, 8, 9, 10, 11, 13, 14, 15, 16, 18, 19 <223> / Modified base = "2'-O-methyl corresponding nucleotide" <221> Modified base <222> 21 <223> / Modified base = "5'-thio-phosphate thymidine" <221> Modified base <222> 3, 5, 7, 12, 17 <223> / Modified base = "2'-hydroxy corresponding nucleotide" <400> 421 gugugcacuu cgcuucacct t 21 <210> 422 <211> 21 <212> DNA <213> Artificial sequence <223> Artificial sequence description: Sense strand of dsRNA <221> Modified base <222> 1 <223> / Modified base = "Nucleotide: lacking 5'-phosphate group" <221> Modified base <222> 1, 2, 3, 4, 6, 8, 9, 11, 15, 19 <223> / Modified base = "2'-O-methyl corresponding nucleotide" <221> Modified base <222> 21 <223> / Modified base = "5'-thio-phosphate thymidine" <221> Modified base <222> 5, 7, 10, 12, 13, 14, 16, 17, 18 <223> / Modified base = "2'-hydroxy corresponding nucleotide" <400> 422 gcucguguua caggcgggct t 21 <210> 423 <211> 21 <212> DNA <213> Artificial sequence <223> Description of artificial sequence: sense strand of dsRNA <221> Modified base <222> 1 <223> / Modified base = "nucleotide: lacking 5'-phosphate group" <221> Modified base <222> 1, 2, 3, 5, 6, 7, 8, 10, 11, 14, 16, 17, 18, 19 <223> / Modified base = "2'-O-methyl corresponding nucleotide" <221> Modified base <222> 21 <223> / Modified base = "5'-thio-phosphate thymidine" <221> Modified base <222> 4, 9, 12, 13, 15 <223> / Modified base = "2'-hydroxy corresponding nucleotide" <400> 423 ccuaucuuau caacacuuct t 21 <210> 424 <211> 21 <212> DNA <213> Artificial sequence <223> Description of artificial sequence: sense strand of dsRNA <221> Modified base <222> 1 <223> / Modified base = "nucleotide: lacking 5'-phosphate group" <221> Modified base <222> 1, 2, 3, 4, 7, 8, 10, 11, 13, 15, 16, 18 <223> / Modified base = "2'-O-methyl corresponding nucleotide" <221> Modified base <222> 21 <223> / Modified base = "5'-thio-phosphate thymidine" <221> Modified base <222> 5, 6, 9, 12, 14, 17, 19 <223> / Modified base = "2'-hydroxy corresponding nucleotide" <400> 424 ucucaaucgc cgcgucgcat t 21 <210> 425 <211> 21 <212> DNA <213> Artificial sequence <223> Artificial sequence description: Sense strand of dsRNA <221> Modified base <222> 1 <223> / Modified base = "Nucleotide: lacking 5'-phosphate group" <221> Modified base <222> 1, 2, 3, 4, 6, 7, 8, 10, 12, 13, 14, 15, 16, 17, 18 <223> / Modified base = "2'-O-methyl corresponding nucleotide" <221> Modified base <222> 21 <223> / Modified base = "5'-thio-phosphate thymidine" <221> Modified base <222> 5, 9, 11, 19 <223> / Modified base = "2'-hydroxy corresponding nucleotide" <400> 425 gcccgucugu gccuucucat t 21 <210> 426 <211> 21 <212> DNA <213> Artificial sequence <223> Artificial sequence description: Sense strand of dsRNA <221> Modified base <222> 1 <223> / Modified base = "Nucleotide: lacking 5'-phosphate group" <221> Modified base <222> 1, 2, 4, 5, 6, 7, 9, 10, 13, 15, 16, 17, 18, 19 <223> / Modified base = "2'-O-methyl corresponding nucleotide" <221> Modified base <222> 21 <223> / Modified base = "5'-thio-phosphate thymidine" <221> Modified base <222> 3, 8, 11, 12, 14 <223> / Modified base = "2'-hydroxy corresponding nucleotide" <400> 426 cuaucuuauc aacacuucct t 21 <210> 427 <211> 21 <212> DNA <213> Artificial sequence <223> Artificial sequence description: Sense strand of dsRNA <221> Modified base <222> 1 <223> / Modified base = "Nucleotide: lacking 5'-phosphate group" <221> Modified base <222> 1, 2, 4, 5, 7, 8, 9, 11, 12, 13, 15, 16, 17, 18, 19 <223> / Modified base = "2'-O-methyl corresponding nucleotide" <221> Modified base <222> 21 <223> / Modified base = "5'-thio-phosphate thymidine" <221> Modified base <222> 3, 6, 10, 14 <223> / Modified base = "2'-hydroxy corresponding nucleotide" <400> 427 auguugcccg uuuguccuct t 21 <210> 428 <211> 21 <212> DNA <213> Artificial sequence <223> Artificial sequence description: Sense strand of dsRNA <221> Modified base <222> 1 <223> / Modified base = "Nucleotide: lacking 5'-phosphate group" <221> Modified base <222> 1, 2, 4, 6, 7, 9, 10, 11, 13, 14, 15, 17, 18, 19 <223> / Modified base = "2'-O-methyl corresponding nucleotide" <221> Modified base <222> 21 <223> / Modified base = "5'-thio-phosphate thymidine" <221> Modified base <222> 3, 5, 8, 12, 16 <223> / Modified base = "2'-hydroxy corresponding nucleotide" <400> 428 guauguugcc cguuugucct t 21 <210> 429 <211> 21 <212> DNA <213> Artificial sequence <223> Artificial sequence description: sense strand of dsRNA <221> Modified base <222> 1 <223> / Modified base = "Nucleotide: lacking 5'-phosphate group" <221> Modified base <222> 1, 2, 3, 4, 6, 7, 8, 9, 11, 12, 13, 14, 15, 17, 19 <223> / Modified base = "2'-O-methyl corresponding nucleotide" <221> Modified base <222> 21 <223> / Modified base = "5'-thio-phosphate thymidine" <221> Modified base <222> 5, 10, 16, 18 <223> / Modified base = "2'-hydroxy corresponding nucleotide" <400> 429 cuucgcuuca ccucugcact t 21 <210> 430 <211> 21 <212> DNA <213> Artificial sequence <223> Description of artificial sequence: sense strand of dsRNA <221> Modified base <222> 1 <223> / Modified base = "nucleotide: lacking 5'-phosphate group" <221> Modified base <222> 1, 2, 3, 5, 7, 8, 9, 10, 12, 13, 14, 15, 17, 18, 19 <223> / Modified base = "2'-O-methyl corresponding nucleotide" <221> Modified base <222> 21 <223> / Modified base = "5'-thio-phosphate thymidine" <221> Modified base <222> 4, 6, 11, 16 <223> / Modified base = "2'-hydroxy corresponding nucleotide" <400> 430 ugugcacuuc gcuucaccut t 21 <210> 431 <211> 21 <212> DNA <213> Artificial sequence <223> Description of artificial sequence: sense strand of dsRNA <221> Modified base <222> 1 <223> / Modified base = "nucleotide: lacking 5'-phosphate group" <221> Modified base <222> 1, 2, 3, 8, 9, 10, 12, 15, 16, 17, 18 <223> / Modified base = "2'-O-methyl corresponding nucleotide" <221> Modified base <222> 21 <223> / Modified base = "5'-thiolated thymidine" <221> Modified base <222> 4, 5, 6, 7, 11, 13, 14, 19 <223> / Modified base = "2'-hydroxy corresponding nucleotide" <400> 431 gccaaaauuc gcagucccgt t 21 <210> 432 <211> 21 <212> DNA <213> Artificial sequence <223> Description of artificial sequence: Sense strand of dsRNA <221> Modified base <222> 1 <223> / Modified base = "Nucleotide: lacking 5'-phosphate group" <221> Modified base <222> 1, 2, 3, 5, 6, 7, 9, 11, 12, 14, 18 <223> / Modified base = "2'-O-methyl corresponding nucleotide" <221> Modified base <222> 21 <223> / Modified base = "5'-thiolated thymidine" <221> Modified base <222> 4, 8, 10, 13, 15, 16, 17, 19 <223> / Modified base = "2'-hydroxy corresponding nucleotide" <400> 432 ccugcucgug uuacaggcgt t 21 <210> 433 <211> 21 <212> DNA <213> Artificial sequence <223> Description of artificial sequence: Sense strand of dsRNA <221> Modified base <222> 1 <223> / Modified base = "Nucleotide: lacking 5'-phosphate group" <221> Modified base <222> 1, 2, 6, 8, 12, 13, 14, 16, 18, 19 <223> / Modified base = "2'-O-methyl corresponding nucleotide" <221> Modified base <222> 21 <223> / Modified base = "5'-thiophosphate thymidine" <221> Modified base <222> 3, 4, 5, 7, 9, 10, 11, 15, 17 <223> / Modified base = "2'-hydroxy corresponding nucleotide" <400> 433 uggagugugg auucgcacut t 21 <210> 434 <211> 21 <212> DNA <213> Artificial sequence <223> Artificial sequence description: Sense strand of dsRNA <221> Modified base <222> 1 <223> / Modified base = "Nucleotide: lacking 5'-phosphate group" <221> Modified base <222> 1, 2, 3, 6, 7, 10, 11, 12, 13, 18 <223> / Modified base = "2'-O-methyl corresponding nucleotide" <221> Modified base <222> 21 <223> / Modified base = "5'-thiophosphate thymidine" <221> Modified base <222> 4, 5, 8, 9, 14, 15, 16, 17, 19 <223> / Modified base = "2'-hydroxy corresponding nucleotide" <400> 434 aacgaccgac cuugaggcat t 21 <210> 435 <211> 21 <212> DNA <213> Artificial sequence <223> Description of artificial sequence: sense strand of dsRNA <221> Modified base <222> 1 <223> / Modified base = "Nucleotide: lacking 5'-phosphate group" <221> Modified base <222> 1, 2, 7, 8, 9, 13, 14, 15, 17 <223> / Modified base = "2'-O-methyl corresponding nucleotide" <221> Modified base <222> 21 <223> / Modified base = "5'-thiophosphate thymidine" <221> Modified base <222> 3, 4, 5, 6, 10, 11, 12, 16, 18, 19 <223> / Modified base = "2'-hydroxy corresponding nucleotide" <400> 435 acagagucua gacucguggt t 21 <210> 436 <211> 21 <212> DNA <213> Artificial sequence <223> Description of artificial sequence: sense strand of dsRNA <221> Modified base <222> 1 <223> / Modified base = "Nucleotide: lacking 5'-phosphate group" <221> Modified base <222> 1, 2, 3, 4, 6, 7, 9, 11, 12, 14 <223> / Modified base = "2'-O-methyl corresponding nucleotide" <221> Modified base <222> 21 <223> / Modified base = "5'-thiophosphate thymidine" <221> Modified base <222> 5, 8, 10, 13, 15, 16, 17, 18, 19 <223> / Modified base = "2'-hydroxy corresponding nucleotide" <400> 436 aaucgccgcg ucgcagaagt t 21 <210> 437 <211> 21 <212> DNA <213> Artificial sequence <223> Artificial sequence description: Sense strand of dsRNA <221> Modified base <222> 1 <223> / Modified base = "Nucleotide: lacking 5'-phosphate group" <221> Modified base <222> 1, 2, 3, 5, 7, 8, 10, 11, 12, 14, 15, 16, 18, 19 <223> / Modified base = "2'-O-methyl corresponding nucleotide" <221> Modified base <222> 21 <223> / Modified base = "5'-thiophosphate thymidine" <221> Modified base <222> 4, 6, 9, 13, 17 <223> / Modified base = "2'-hydroxy corresponding nucleotide" <400> 437 gguauguugc ccguuuguct t 21 <210> 438 <211> 21 <212> DNA <213> Artificial sequence <223> Artificial sequence description: Sense strand of dsRNA <221> Modified base <222> 1 <223> / Modified base = "Nucleotide: lacking 5'-phosphate group" <221> Modified base <222> 1, 2, 3, 6, 7, 8, 10, 12, 13, 15 <223> / Modified base = "2'-O-methyl corresponding nucleotide" <221> Modified base <222> 21 <223> / Modified base = "5'-thio-phosphate thymidine" <221> Modified base <222> 4, 5, 9, 11, 14, 16, 17, 18, 19 <223> / Modified base = "2'-hydroxy corresponding nucleotide" <400> 438 gccgauccau acugcggaat t 21 <210> 439 <211> 21 <212> DNA <213> Artificial sequence <223> Artificial sequence description: Sense strand of dsRNA <221> Modified base <222> 1 <223> / Modified base = "Nucleotide: lacking 5'-phosphate group" <221> Modified base <222> 1, 2, 3, 4, 5, 7, 8, 9, 10, 12, 13, 16, 18, 19 <223> / Modified base = "2'-O-methyl corresponding nucleotide" <221> Modified base <222> 21 <223> / Modified base = "5'-thio-phosphate thymidine" <221> Modified base <222> 6, 11, 14, 15, 17 <223> / Modified base = "2'-hydroxy corresponding nucleotide" <400> 439 gcccuaucuu aucaacacut t 21 <210> 440 <211> 21 <212> DNA <213> Artificial sequence <223> Description of artificial sequence: Sense strand of dsRNA <221> Modified base <222> 1 <223> / Modified base = "Nucleotide: lacking 5'-phosphate group" <221> Modified base <222> 1, 2, 3, 4, 5, 7, 8, 11, 13, 14, 16, 17, 18 <223> / Modified base = "2'-O-methylated corresponding nucleotide" <221> Modified base <222> 21 <223> / Modified base = "5'-thio-phosphate thymidine" <221> Modified base <222> 6, 9, 10, 12, 15, 19 <223> / Modified base = "2'-hydroxylated corresponding nucleotide" <400> 440 aguuuacuag ugccauuugt t 21 <210> 441 <211> 21 <212> DNA <213> Artificial sequence <223> Description of artificial sequence: Sense strand of dsRNA <221> Modified base <222> 1 <223> / Modified base = "Nucleotide: lacking 5'-phosphate group" <221> Modified base <222> 1, 2, 3, 4, 7, 10, 11, 14, 15, 16, 17 <223> / Modified base = "2'-O-methylated corresponding nucleotide" <221> Modified base <222> 21 <223> / Modified base = "5'-thio-phosphate thymidine" <221> Modified base <222> 5, 6, 8, 9, 12, 13, 18, 19 <223> / Modified base = "2'-hydroxy corresponding nucleotide" <400> 441 ugucaacgac cgaccuugat t 21 <210> 442 <211> 21 <212> DNA <213> Artificial sequence <223> Artificial sequence description: Sense strand of dsRNA <221> Modified base <222> 1 <223> / Modified base = "Nucleotide: lacking 5'-phosphate group" <221> Modified base <222> 1, 2, 3, 4, 5, 6, 7, 8, 9, 12, 13, 14, 15, 16, 17 <223> / Modified base = "2'-O-methyl corresponding nucleotide" <221> Modified base <222> 21 <223> / Modified base = "5'-thio-phosphate thymidine" <221> Modified base <222> 10, 11, 18, 19 <223> / Modified base = "2'-hydroxy corresponding nucleotide" <400> 442 acuucucuca auuuucuagt t 21 <210> 443 <211> 21 <212> DNA <213> Artificial sequence <223> Artificial sequence description: Sense strand of dsRNA <221> Modified base <222> 1 <223> / Modified base = "Nucleotide: lacking 5'-phosphate group" <221> Modified base <222> 1, 2, 4, 9, 11, 12, 13, 14, 15, 16, 18, 19 <223> / Modified base = "2'-O-methyl corresponding nucleotide" <221> Modified base <222> 21 <223> / Modified base = "5'-thio-phosphate thymidine" <221> Modified base <222> 3, 5, 6, 7, 8, 10, 17 <223> / Modified base = "2'-hydroxy corresponding nucleotide" <400> 443 gcgcgggacg uccuuuguct t 21 <210> 444 <211> 21 <212> DNA <213> Artificial sequence <223> Artificial sequence description: Sense strand of dsRNA <221> Modified base <222> 1 <223> / Modified base = "Nucleotide: lacking 5'-phosphate group" <221> Modified base <222> 1, 2, 3, 7, 8, 9, 11, 14, 18, 19 <223> / Modified base = "2'-O-methyl corresponding nucleotide" <221> Modified base <222> 21 <223> / Modified base = "5'-thio-phosphate thymidine" <221> Modified base <222> 4, 5, 6, 10, 12, 13, 15, 16, 17 <223> / Modified base = "2'-hydroxy corresponding nucleotide" <400> 444 ucuagacucg ugguggacut t 21 <210> 445 <211> 21 <212> DNA <213> Artificial sequence <223> Artificial sequence description: Sense strand of dsRNA <221> Modified base <222> 1 <223> / Modified base = "Nucleotide: lacking 5'-phosphate group" <221> Modified base <222> 1, 2, 3, 4, 5, 7, 9, 10, 12, 17, 18, 19 <223> / Modified base = "2'-O-methyl corresponding nucleotide" <221> Modified base <222> 21 <223> / Modified base = "5'-thiophosphate thymidine" <221> Modified base <222> 6, 8, 11, 13, 14, 15, 16 <223> / Modified base = "2'-hydroxy corresponding nucleotide" <400> 445 gauccauacu gcggaacuct t 21 <210> 446 <211> 21 <212> DNA <213> Artificial sequence <223> Artificial sequence description: Sense strand of dsRNA <221> Modified base <222> 1 <223> / Modified base = "Nucleotide: lacking 5'-phosphate group" <221> Modified base <222> 1, 2, 3, 4, 6, 7, 10, 11, 12, 14, 16, 17, 19 <223> / Modified base = "2'-O-methyl corresponding nucleotide" <221> Modified base <222> 21 <223> / Modified base = "5'-thiophosphate thymidine" <221> Modified base <222> 5, 8, 9, 13, 15, 18 <223> / Modified base = "2'-hydroxy corresponding nucleotide" <400> 446 cucugccgau ccauacugct t 21 <210> 447 <211> 21 <212> DNA <213> Artificial sequence <223> Description of artificial sequence: sense strand of dsRNA <221> Modified base <222> 1 <223> / Modified base = "Nucleotide: lacking 5'-phosphate group" <221> Modified base <222> 1, 2, 3, 5, 6, 9, 10, 11, 13, 15, 16, 18 <223> / Modified base = "2'-O-methyl corresponding nucleotide" <221> Modified base <222> 21 <223> / Modified base = "5'-thio-phosphate thymidine" <221> Modified base <222> 4, 7, 8, 12, 14, 17, 19 <223> / Modified base = "2'-hydroxy corresponding nucleotide" <400> 447 ucugccgauc cauacugcgt t 21 <210> 448 <211> 21 <212> DNA <213> Artificial sequence <223> Description of artificial sequence: sense strand of dsRNA <221> Modified base <222> 1 <223> / Modified base = "Nucleotide: lacking 5'-phosphate group" <221> Modified base <222> 1, 2, 3, 4, 5, 7, 8, 11, 12, 13, 15, 17, 18 <223> / Modified base = "2'-O-methyl corresponding nucleotide" <221> Modified base <222> 21 <223> / Modified base = "5'-thiophosphate thymidine" <221> Modified base <222> 6, 9, 10, 14, 16, 19 <223> / Modified base = "2'-hydroxy corresponding nucleotide" <400> 448 ccucugccga uccauacugt t 21 <210> 449 <211> 21 <212> DNA <213> Artificial sequence <223> Artificial sequence description: Sense strand of dsRNA <221> Modified base <222> 1 <223> / Modified base = "Nucleotide: lacking 5'-phosphate group" <221> Modified base <222> 1, 2, 4, 5, 6, 7, 8, 9, 10, 11, 12, 14, 16, 19 <223> / Modified base = "2'-O-methyl corresponding nucleotide" <221> Modified base <222> 21 <223> / Modified base = "5'-thiophosphate thymidine" <221> Modified base <222> 3, 13, 15, 17, 18 <223> / Modified base = "2'-hydroxy corresponding nucleotide" <400> 449 gcaccucucu uuacgcggut t 21 <210> 450 <211> 21 <212> DNA <213> Artificial sequence <223> Artificial sequence description: Sense strand of dsRNA <221> Modified base <222> 1 <223> / Modified base = "Nucleotide: lacking 5'-phosphate group" <221> Modified base <222> 1, 2, 6, 7, 8, 9, 10, 11, 12, 14, 15, 16, 17, 19 <223> / Modified base = "2'-O-methyl corresponding nucleotide" <221> Modified base <222> 21 <223> / Modified base = "5'-thiophosphate thymidine" <221> Modified base <222> 3, 4, 5, 13, 18 <223> / Modified base = "2'-hydroxy corresponding nucleotide" <400> 450 aagaacuccc ucgccucgct t 21 <210> 451 <211> 21 <212> DNA <213> Artificial sequence <223> Description of artificial sequence: Sense strand of dsRNA <221> Modified base <222> 1 <223> / Modified base = "Nucleotide: lacking 5'-phosphate group" <221> Modified base <222> 1, 2, 4, 5, 6, 7, 8, 9, 10, 12, 13, 14, 15, 17 <223> / Modified base = "2'-O-methyl corresponding nucleotide" <221> Modified base <222> 21 <223> / Modified base = "5'-thiophosphate thymidine" <221> Modified base <222> 3, 11, 16, 18, 19 <223> / Modified base = "2'-hydroxy corresponding nucleotide" <400> 451 gaacucccuc gccucgcagt t 21 <210> 452 <211> 21 <212> DNA <213> Artificial sequence <223> Description of artificial sequence: sense strand of dsRNA <221> Modified base <222> 1 <223> / Modified base = "nucleotide: lacking 5'-phosphate group" <221> Modified base <222> 1, 2, 3, 4, 5, 6, 9, 10, 11, 12, 13, 14, 19 <223> / Modified base = "2'-O-methylated corresponding nucleotide" <221> Modified base <222> 21 <223> / Modified base = "5'-thiophosphate thymidine" <221> Modified base <222> 7, 8, 15, 16, 17, 18 <223> / Modified base = "2'-hydroxylated corresponding nucleotide" <400> 452 ucucucaauu uucuagggct t 21 <210> 453 <211> 21 <212> DNA <213> Artificial sequence <223> Description of artificial sequence: sense strand of dsRNA <221> Modified base <222> 1 <223> / Modified base = "nucleotide: lacking 5'-phosphate group" <221> Modified base <222> 1, 2, 4, 6, 8, 9, 10, 11, 12, 13, 14, 15, 16, 18 <223> / Modified base = "2'-O-methylated corresponding nucleotide" <221> Modified base <222> 21 <223> / Modified base = "5'-thiophosphate thymidine" <221> Modified base <222> 3, 5, 7, 17, 19 <223> / Modified base = "2'-hydroxy corresponding nucleotide" <400> 453 gggcgcaccu cucuuuacgt t 21 <210> 454 <211> 21 <212> DNA <213> Artificial sequence <223> Artificial sequence description: Sense strand of dsRNA <221> Modified base <222> 1 <223> / Modified base = "Nucleotide: lacking 5'-phosphate group" <221> Modified base <222> 1, 2, 5, 6, 7, 9, 11, 12, 14, 19 <223> / Modified base = "2'-O-methyl corresponding nucleotide" <221> Modified base <222> 21 <223> / Modified base = "5'-thio-phosphate thymidine" <221> Modified base <222> 3, 4, 8, 10, 13, 15, 16, 17, 18 <223> / Modified base = "2'-hydroxy corresponding nucleotide" <400> 454 ccgauccaua cugcggaact t 21 <210> 455 <211> 21 <212> DNA <213> Artificial sequence <223> Artificial sequence description: Sense strand of dsRNA <221> Modified base <222> 1 <223> / Modified base = "Nucleotide: lacking 5'-phosphate group" <221> Modified base <222> 1, 2, 3, 4, 5, 6, 7, 8, 9, 11, 12, 13, 14, 16 <223> / Modified base = "2'-O-methyl corresponding nucleotide" <221> Modified base <222> 21 <223> / Modified base = "5'-thiophosphate thymidine" <221> Modified base <222> 10, 15, 17, 18, 19 <223> / Modified base = "2'-hydroxy corresponding nucleotide" <400> 455 aacucccucg ccuc...

Claims

1. A double-stranded RNA molecule capable of inhibiting the expression of hepatitis B virus gene, wherein the double-stranded RNA molecule comprises a sense strand and an antisense strand containing the sequence described in SEQ ID NO: 2, and the antisense strand is complementary to the sense strand, wherein the length of the double-stranded RNA molecule is 19-25 nucleotides.

2. A double-stranded ribonucleic acid molecule as described in claim 1, wherein the antisense strand comprises nucleotides 2-18 of SEQ ID NO:

158. The double-stranded RNA molecule of claim 1 , comprising at least one modified nucleotide.

4. A double-stranded ribonucleic acid molecule as described in claim 3, wherein the at least one modified nucleotide is selected from: 2'-O-methyl modified nucleotides, nucleotides containing 5'-thiophosphate groups, terminal nucleotides linked to cholesterol derivatives, terminal nucleotides linked to dodecanoic acid didecylamide groups, 2'-deoxy-2'-fluoro modified nucleotides, 2'-deoxy-modified nucleotides, locked nucleotides, abasic nucleotides, deoxythymidine, inverted deoxythymidine, 2'-amino-modified nucleotides, 2'-alkyl-modified nucleotides, morpholino nucleotides, phosphoramidates and nucleotides containing non-natural bases.

5. The double-stranded ribonucleic acid molecule of claim 4, comprising 2'-O-methyl modified nucleotides, nucleotides comprising 5'-phosphorothioate groups and deoxythymidine. The double-stranded RNA molecule of claim 1 , wherein the sense strand comprises a 3′-overhang of 1-2 nucleotides.

7. The double-stranded ribonucleic acid molecule of claim 1, wherein the antisense strand comprises a 3'-overhang of 1-2 nucleotides. The double-stranded RNA molecule of claim 1 , wherein the double-stranded RNA molecule is conjugated to a ligand.

9. The double-stranded ribonucleic acid molecule of claim 8, wherein the ligand is a sugar cluster.

10. The double-stranded ribonucleic acid molecule of claim 8, wherein the ligand comprises galactose.

11. A pharmaceutical composition comprising the double-stranded ribonucleic acid molecule according to any one of claims 1 to 10 and a pharmaceutically acceptable carrier.

12. Use of the double-stranded RNA molecule according to any one of claims 1 to 10 in the preparation of a medicament for treating hepatitis B virus infection.

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