小型干擾核糖核酸已成為在哺乳動物系統中研究基因功能最強力的基因沉默試劑。然而，一些實驗證據顯示，只有一小部分的siRNAs才能引發高度有效的核糖核酸干擾。此外，siRNAs的效力只能藉由實驗上抑制標的基因的表現來測量。為了使有效siRNA的篩選更健全和經濟，我們已經發展出一個簡易、可定量且不需要cDNA的siRNA確認系統。此系統包含了一段含有RNAi標的序列的短合成DNA片段，以及標的報告載體和驅動siRNA表現載體。為了產生標的報告載體，我們把合成的短DNA片段取代cDNA融合在報告基因的5’端非轉譯區、3’端非轉譯區或是二個報告基因中間而不影響報告基因的活性。此DNA片段又可同時放入老鼠U6及人類H1二個相向的RNA polymerase III（pol III） 啟動子之間，以各別驅動siRNA的正義及反義股之表現。siRNA的效力只能藉由專一性地抑制標的報告基因（EGFP或firefly luciferase）的表現而得知。藉著直接針對B型肝炎表面抗原（HBsAg）及腫瘤抑制蛋白p53，且已經被確認為有效的siRNA，證明了我們所建立的這個siRNA確認系統可以有效率且忠誠地反應出siRNA的真實效力。我們又更進一步地利用此系統篩選出高度有效的siRNA來抑制老鼠MMP-7。既然只需要一段容易獲得的合成短DNA片段就可以同時構築至標的報告載體和驅動siRNA表現載體，這個新的系統不但加速了在哺乳動物系統中大規模功能失去（loss-of-function）的基因篩選，並且完全提供了一個較良善的方法去篩選及認定可以成為治療試劑的有效siRNA。另一方面，我們分析了H1啟動子雙向調節性的特質。我們利用EGFP及firefly luciferase基因驗證H1啟動子是一個雙向啟動子，並應用其雙向調節的特性同時表現抗藥基因與shLuc來篩選穩定細胞株。

　Small interfering RNAs (siRNA) have become the most powerful gene silencing reagents for studying gene functions in mammalian systems. However, some experimental evidences have already shown that only a limited number of siRNAs can induce highly effective RNA interference (RNAi) in mammalian cells. Moreover, the efficacy of siRNAs can only be experimentally measured based on suppression of the target gene expression. To achieve the functional screening for effective siRNAs more robustly and cost-effectively, we have developed a reliable and quantitative reporter-based siRNA validation system that eliminated the need for cDNA clones. This system consisted of a short synthetic DNA fragment including an RNAi targeting sequence of interest and two expression vectors for targeting reporter as well as triggering siRNA expression. In order to generate the targeting reporter vector for this system, the short synthetic DNA fragment, instead of cDNA, was fused with a reporter gene at the 5’-or 3’-untranslated region (5’-or 3’-UTR) or inserted within the reporter gene without interrupting its activity. Simultaneously, this DNA fragment was also cloned into the triggering siRNA expression vector, pDual, which contained two convergent RNA polymerase III (Pol III) promoters, mouse U6 and human H1, to drive expression of the sense and antisense strands of siRNA, respectively. The efficacy of the siRNAs is determined by their abilities to inhibit expression of the targeting reporter gene with easily quantified readouts including enhanced green fluorescence protein (EGFP) or firefly luciferase. By using fully analyzed siRNAs directed against human hepatitis B virus (HBV) surface antigen (HBsAg) or tumor suppressor protein p53 (Trp53) gene expression, we have demonstrated that this reporter-based siRNA validation system could effectively and faithfully report the efficacy of the corresponding siRNAs in a sequence-specific manner. In addition, we have further identified the potent siRNAs for silencing mouse MMP-7 gene expression by using this system. Furthermore, H1 promoter is often used to express siRNA or shRNA, and it has a bidirectional character. We have confirmed H1 promoter was a bidirectional promoter by using EGFP as well as firefly luciferase genes. In addition, we have selected stable cell lines expressing Bsd and shLuc by applying H1 promoter.

Abbas-Terki, T., Blanco-Bose, W., Deglon, N., Pralong, W. and Aebischer, P. Lentiviral-mediated RNA interference. Hum. Gene Ther. 13, 2197-2201 (2002).
Adelman, Z. N., Sanchez-Vargas, I., Travanty, E. A., Carlson, J. O., Beaty, B. J., Blair, C. D. and Olson, K. E. RNA silencing of dengue virus type 2 replication in transformed C6/36 mosquito cells transcribing an inverted-repeat RNA derived from the virus genome. J. Virol. 76, 12925-12933 (2002).
Amé, J. C., Rolli, V., Schreiber, V., Niedergang, C., Apiou, F., Decker, P., Muller, S., Höger, T., Ménissier-de Murcia, J., and de Murcia, G. PARP-2, A Novel Mammalian DNA Damage-dependent Poly (ADP-ribose) Polymerase J. Biol. Chem. 274, 17860-17868 (1999).
Amé, J. C., Schreiber, V., Fraulob, V., Dolle, P., de Murcia, G. and Niedergang, C. P. A bidirectional promoter connects the poly(ADP-ribose) polymerase 2 (PARP-2) gene to the gene for RNase P RNA. structure and expression of the mouse PARP-2 gene. J. Biol. Chem. 276, 11092-11099 (2001).
Ashrafi, K., Chang, F. Y., Watts, J. L., Fraser, A. G., Kamath, R. S., Ahringer, J. and Ruvkun, G. Genome-wide RNAi analysis of Caenorhabditis elegans fat regulatory genes. Nature 421, 268-272 (2003) .
Baer, M., Nilsen, T. W., Costigan, C. and Altman, S. Structure and transcription of a human gene for H1 RNA, the RNA component of human RNase P. Nucleic Acids Res. 18, 97-103 (1990).
Bai, J., Ramos, R. L., Ackman, J. B., Thomas, A. M., Lee, R. V. and LoTurco, J. J. RNAi reveals doublecortin is required for radial migration in rat neocortex. Nat. Neurosci. 6, 1277-1283 (2003).
Banerjee, D. and Slack, F. Control of developmental timing by small temporal RNAs: a paradigm for RNA-mediated regulation of gene expression. Bioessays 24, 119-129 (2002).
Bartel, D.P. MicroRNAs:genomics , biogenesis, mechanism, and function. Cell 116, 218-297 (2004).
Barton, G. M. and Medzhitov, R. Retroviral delivery of small interfering RNA into primary cells. Proc. Natl. Acad. Sci. U S A. 99, 14943-14945 (2002).
Basset, P., Bellocq, J. P., Wolf, C., Stoll, I., Hutin, P., Limacher, J. M., Podhajcer, O. L., Chenard, M. P., Rio, M. C. and Chambon, P. A novel metalloproteinase gene specifically expressed in stromal cells of breast carcinomas. Nature 348, 699-704 (1990).
Berns, K., Hijmans, E. M., Mullenders, J., Brummelkamp, T. R., Velds, A., Heimerikx, M., Kerkhoven, R. M., Madiredjo, M., Nijkamp, W., Weigelt, B., Agami, R., Ge, W., Cavet, G., Linsley, P. S., Beijersbergen, R. L. and Bernards, R. A large-scale RNAi screen in human cells identifies new components of the p53 pathway. Nature 428, 431-437 (2004).
Bernstein, E., Caudy, A. A., Hammond, S. M. and Hannon, G. J. Role for a bidentate ribonuclease in the initiation step of RNA interference. Nature 409, 363-366 (2001).
Bitko, V., Musiyenko, A., Shulyayeva, O., and Barik, S. Inhibition of respiratory viruses by nasally administered siRNA. Nat. Med. 11, 50-55 (2005).
Boden, D., Pusch, O., Silbermann, R., Lee, F., Tucker, L. and Ramratnam, B. Enhanced gene silencing of HIV-1 specific siRNA using microRNA designed hairpins. Nucleic Acids Res. 32, 1154-1158 (2004).
Bohula, E. A., Salisbury, A. J., Sohail, M., Playford, M. P., Riedemann, J., Southern, E. M. and Macaulay, V. M. The efficacy of small interfering RNAs targeted to the type 1 insulin-like growth factor receptor (IGF1R) is influenced by secondary structure in the IGF1R transcript. J. Biol. Chem. 278, 15991-15997 (2003).
Brummelkamp, T. R., Bernards, R. and Agami, R. A system for stable expression of short interfering RNAs in mammalian cells. Science 296, 550-553 (2002).
Brummelkamp, T. R., Bernards, R. and Agami, R. Stable suppression of tumorigenicity by virus-mediated RNA interference. Cancer Cell 2, 243-247 (2002).
Bullock, A. N. and Fersht, A. R. Rescuing the function of mutant p53. Nat. Rev. Cancer 1, 68-76 (2001).
Calegari, F., Haubensak, W., Yang, D., Huttner, W. B. and Buchholz, F. Tissue-specific RNA interference in postimplantation mouse embryos with endoribonuclease-prepared short interfering RNA. Proc. Natl. Acad. Sci. U S A. 99, 14236-14240 (2002).
Calin, G. A., Dumitru, C. D., Shimizu, M., Bichi, R., Zupo, S., Noch, E., Aldler, H., Rattan, S., Keating, M., Rai, K., Rassenti, L., Kipps, T., Negrini, M., Bullrich, F. and Croce, C. M. Frequent deletions and down-regulation of micro- RNA genes miR15 and miR16 at 13q14 in chronic lymphocytic leukemia. Proc. Natl. Acad. Sci. U S A. 99, 15524-15529 (2002).
Calin, G. A., Liu, C. G., Sevignani, C., Ferracin, M., Felli, N., Dumitru, C. D., Shimizu, M., Cimmino, A., Zupo, S., Dono, M., Dell'Aquila, M. L., Alder, H., Rassenti, L., Kipps, T. J., Bullrich, F., Negrini, M. and Croce, C. M. MicroRNA profiling reveals distinct signatures in B cell chronic lymphocytic leukemias. Proc. Natl. Acad. Sci. U S A. 101, 11755-11760 (2004).
Caplen, N. J., Zheng, Z., Falgout, B. and Morgan, R. A. Inhibition of viral gene expression and replication in mosquito cells by dsRNA-triggered RNA interference. Mol. Ther. 6, 243-251 (2002).
Carbon, P., Murgo, S., Ebel, J. P., Krol, A., Tebb, G. and Mattaj, J. A common octamer motif binding protein is involved in the transcription of U6 snRNA by RNA polymerase III and U2 snRNA by RNA polymerase II. Cell 51, 71-79 (1987).
Chang, S., Johnston, R. J. Jr., Frokjaer-Jensen, C., Lockery, S. and Hobert, O. MicroRNAs act sequentially and assmmetrically to control chemosensory laterality in the nematode. Nature 430, 785-798 (2004).
Chen, Z., Indjeian, V. B., McManus, M., Wang, L. and Dynlacht, B. D. CP110, a cell cycle-dependent CDK substrate, regulates centrosome duplication in human cells. Dev. Cell 3, 339-350 (2002).
Chiu, Y. L., and Rana, T. M. RNAi in human cells：basic structural and functional features of small interfering RNA. Mol. Cell 10, 549-561 (2002).
Chiu, Y. L. and Rana, T. M. siRNA function in RNAi: a chemical modification analysis. RNA 9, 1034-1048 (2003).
Cioca, D. P., Aoki, Y. and Kiyosawa, K. RNA interference is a functional pathway with therapeutic potential in human myeloid leukemia cell lines. Cancer Gene Ther. 10,125-133 (2003).
Clayton, J. RNA interference: the silent treatment. Nature 431, 599-605 (2004).
Coburn, G. A. and Cullen, B. R. Potent and specific inhibition of human immunodeficiency virus type 1 replication by RNA interference. J. Virol. 76, 9225-9231 (2002).
Cogoni, C. and Macino, G. Homology-dependent gene silencing in plants and fungi: a number of variations on the same theme. Curr. Opin. Microbiol. 2, 657-662 (1999).
Dalmay, T., Hamilton, A., Rudd, S., Angell, S. and Baulcombe, D. C. An RNA-dependent RNA polymerase gene in Arabidopsis is required for posttranscriptional gene silencing mediated by a transgene but not by a virus. Cell 101, 543-553 (2000).
Das, G., Henning, D., Wright, D. and Reddy, R. Upstream regulatory elements are necessary and sufficient for transcription of a U6 RNA gene by RNA polymerase III. EMBO J. 7, 503-512 (1988).
Davidson, B. L. and Paulson, H. L. Molecular medicine for the brain: silencing of disease genes with RNA interference. Lancet Neurol. 3, 145-149 (2004).
Debes, J. D., Schmidt, L. J., Huang, H. and Tindall, D. J. p300 mediates androgen-independent transactivation of the androgen receptor by interleukin 6. Cancer Res. 62, 5632-5636 (2002).
Dorsett, Y. and Tuschl, T. siRNAs: applications in functional genomics and potential as therapeutics. Nat. Rev. Drug Discov. 3, 318-329 (2004).
Dostie, J., Mourelatos, Z., Yang, M., Sharma, A. and Dreyfuss, G.. Numerous microRNPs in neuronal cells containing novel microRNAs. RNA 9, 180-186 (2003).
Du, Q., Thonberg, H., Zhang, H. Y., Wahlestedt, C. and Liang, Z. Validating siRNA using a reporter made from synthetic DNA oligonucleotides. Biochem. Biophys. Res. Commun. 325, 243-249 (2004).
Dudek, P. and Picard, D. TROD: T7 RNAi Oligo Designer. Nucleic Acids Res. 32, W121-123 (2004).
Dykxhoorn, D. M., Novina, C. D., and Sharp, P. A. Killing the messenger: short RNAs that silence gene expression. Nat. Rev. Mol. Cell Biol. 4, 457-467 (2003).
Elbashir, S. M., Harborth, J., Lendeckel, W., Yalcin, A., Weber, K. and Tuschl, T. Duplexes of 21-nucleotide RNAs mediate RNA interference in cultured mammalian cells. Nature 411, 494-498 (2001).
Elbashir, S. M., Harborth, J., Weber, K. and Tuschl, T. Analysis of gene function in somatic mammalian cells using small interfering RNAs. Methods 26, 199-213 (2002).
Elbashir, S. M., Martinez, J., Patkaniowska, A., Lendeckel, W. and Tuschl, T. Functional anatomy of siRNAs for mediating efficient RNAi in Drosophila melanogaster embryo lysate. EMBO J. 20, 6877-6888 (2001).
Ge, Q., McManus, M. T., Nguyen, T., Shen, C. H., Sharp, P. A., Eisen, H. N. and Chen, J. RNA interference of influenza virus production by directly targeting mRNA for degradation and indirectly inhibiting all viral RNA transcription. Proc. Natl. Acad. Sci. USA. 100, 2718-2723 (2003).
Gesellchen, V. and Boutros, M. Managing the genome: microRNAs in Drosophila. Differentiation 72, 74-80 (2004).
Giladi, H., Ketzinel-Gilad, M., Rivkin, L., Felig, Y., Nussbaum, O. and Galun, E. Small interfering RNA inhibits hepatitis B virus replication in mice. Mol. Ther. 8, 769-776 (2003).
Gitlin, L., Karelsky, S. and Andino, R. Short interfering RNA confers intracellular antiviral immunity in human cells. Nature 418, 430-434 (2002).
Gregory, R. J. and Shiekhattar, R. MicroRNA biogenesis and cancer. Cancer Res. 65, 3509-3512 (2005).
Grishok, A., Pasquinelli, A. E., Conte, D., Li, N., Parrish, S., Ha, I., Baillie, D. L., Fire, A., Ruvkun, G. and Mello, C. C. Genes and mechanisms related to RNA interference regulate expression of the small temporal RNAs that control C. elegans developmental timing. Cell 106, 23-34 (2001).
Hacein-Bey-Abina, S., Von Kalle, C., Schmidt, M., McCormack, M. P., Wulffraat, N., Leboulch, P., Lim, A., Osborne, C. S., Pawliuk, R., Morillon, E., Sorensen, R., Forster, A., Fraser, P., Cohen, J. I., de Saint Basile, G., Alexander, I., Wintergerst, U., Frebourg, T., Aurias, A., Stoppa-Lyonnet, D., Romana, S., Radford-Weiss, I., Gross, F., Valensi, F., Delabesse, E., Macintyre, E., Sigaux, F., Soulier, J., Leiva, L. E., Wissler, M., Prinz, C., Rabbitts, T. H., Le Deist, F., Fischer, A. and Cavazzana-Calvo, M. LMO2-associated clonal T cell proliferation in two patients after gene therapy for SCID-X1. Science 302, 415-419 (2003).
Hall, J. Opinion: Unravelling the general properties of siRNAs: strength in numbers and lessons from the past. Nat. Rev. Genet. 5, 552-557 (2004).
Hamilton, A. J. and Baulcombe, D. C. A species of small antisense RNA in posttranscriptional gene silencing in plants. Science 286, 950-952 (1999).
Hammond, S. M., Bernstein, E., Beach, D. and Hannon, G. J. An RNA-directed nuclease mediates post-transcriptional gene silencing in Drosophila cells. Nature 404, 293-296 (2000).
Hammond, S. M., Boettcher, S., Caudy, A. A., Kobayashi, R. and Hannon, G. J. Argonaute2, a link between genetic and biochemical analyses of RNAi. Science 293,1146-1150 (2001).
Hannon, G. J., Chubb, A., Maroney, P. A., Hannon, G., Altman, S. and Nilsen, T. W. Multiple cis-acting elements are required for RNA polymerase III transcription of the gene encoding H1 RNA, the RNA component of human RNase P. J. Biol. Chem. 266, 22796–22799 (1991).
Hannon, G. J. and Rossi, J. J. Unlocking the potential of the human genome with RNA interference. Nature 431, 371-378 (2004).
Harborth, J., Elbashir, S. M., Bechert, K., Tuschl, T. and Weber, K. Identification of essential genes in cultured mammalian cells using small interfering RNAs. J. Cell Sci. 114, 4557-4565 (2001).
Harborth, J., Elbashir, S. M., Vandenburgh, K., Manninga, H., Scaringe, S. A., Weber, K. and Tuschl, T. Sequence, chemical, and structural variation of small interfering RNAs and short hairpin RNAs and the effect on mammalian gene silencing. Antisense Nucleic Acid Drug Dev. 13, 83-105 (2003).
Harper, S. Q., Staber, P. D., He, X., Eliason, S. L., Martins, I. H., Mao, Q., Yang, L., Kotin, R. M., Paulson, H. L. and Davidson, B. L. RNA interference improves motor and neuropathological abnormalities in a Huntington's disease mouse model. Proc. Natl. Acad. Sci. U S A. 102, 5820-5825 (2005).
Hashimoto, K., Kihira, Y., Matuo, Y. and Usui, T. Expression of matrix metalloproteinase-7 and tissue inhibitor of metalloproteinase-1 in human prostate. J. Urol. 160, 1872-1876 (1998).
He, L., Thomson, J. M., Hemann, M. T., Hernando-Monge, E., Mu, D., Goodson, S., Powers, S., Cordon-Cardo, C., Lowe, S. W., Hannon, G. J. and Hammond, S. M. A microRNA polycistron as a potential human oncogene. Nature 435, 828-833 (2005).
Heale, B. S. E., Soifer, H. S., Bowers, C. and Rossi, J. J. siRNA target site secondary structure predictions using local stable substructures. Nucleic Acids Res. 33, e30 (2005) .
Hemann, M. T., Fridman, J. S., Zilfou, J. T., Hernando, E., Paddison, P. J., Cordon-Cardo, C., Hannon, G. J., Lowe. and S. W. An epi-allelic series of p53 hypomorphs created by stable RNAi produces distinct tumor phenotypes in vivo. Nat. Genet. 33, 396-400 (2003).
Henry, R. W., Sadowski, C. L., Kobayashi, R. and Hernandez, N. A TBP–TAF complex required for transcription of human snRNA genes by RNA polymerases II and III. Nature 374, 653–657 (1995).
Henschel, A., Buchholz, F. and Habermann, B. DEQOR: a web-based tool for the design and quality control of siRNAs. Nucleic Acids Res. 32, W113-120 (2004).
Hernandez, N., Lobo, S., Lister, J., Ifill, S., Ratnasabapathy, R., Sheldon, M. and Johal, L. Transcription of the human U2 and U6 RNA genes. Mol. Biol. Rep. 14, 167 (1990).
Herr, W. Oct-1 and Oct-2: differential transcriptional regulation by proteins that bind to the same DNA sequence. Transcriptional Regulation 1, 1103-1135 (1992).
Herr, W. and Cleary, M. A. The POU domain : versatility in transcriptional regulation by a flexible two-in one DNA-binding domain. Genes Dev. 9, 1679-1693 (1995).
Holen, T., Amarzguioui, M., Wiiger, M. T., Babaie, E. and Prydz, H. Positional effects of short interfering RNAs targeting the human coagulation trigger Tissue Factor. Nucleic Acids Res. 30, 1757-1766 (2002).
Hommel, J. D., Sears, R. M., Georgescu, D., Simmons, D. L. and DiLeone, R. J. Local gene knockdown in the brain using viral-mediated RNA interference. Nat. Med. 9, 1539–1544 (2003).
Hsieh, A. C., Bo, R., Manola, J., Vazquez, F., Bare, O., Khvorova,A., Scaringe, S. and Sellers, W. R. A library of siRNA duplexes targeting the phosphoinositide 3-kinase pathway: determinants of gene silencing for use in cell-based screens. Nucleic Acids Res. 32, 893-901 (2004).
Hu, X., Hipolito, S., Lynn, R., Abraham, V., Ramos, S. and Wong-Staal, F. Relative gene-silencing efficiencies of small interfering RNAs targeting sense and antisense transcripts from the same genetic locus. Nucleic Acids Res. 32, 4609-4617 (2004).
Hutvagner, G. and Zamore, P. D. A microRNA in a multiple-turnover RNAi enzyme complex. Science 297, 2056-2060 (2002).
Ishikawa, T., Ichikawa, Y., Mitsuhashi, M., Momiyama, N., Chishima, T., Tanaka, K., Yamaoka, H., Miyazakic, K., Nagashima, Y., Akitaya, T. and Shimada, H. Matrilysin is associated with progression of colorectal tumor. Cancer Lett. 107, 5-10 (1996).
Jacque, J. M., Triques, K. and Stevenson, M. Modulation of HIV-1 replication by RNA interference. Nature 418, 435-438 (2002).
Jagla, B., Aulner, N., Kelly, P. D., Song, D., Volchuk, A., Zatorski, A., Shum, D., Mayer, T., DE Angelis, D. A., Ouerfelli, O., Rutishauser, U., Rothman, J. E. Sequence characteristics of functional siRNAs. RNA 11, 864-872 (2005).
Jiang, M. and Milner, J. Bcl-2 constitutively suppresses p53-dependent apoptosis in colorectal cancer cells. Genes Dev. 17, 832-837 (2003).
Jin, P., Zarnescu, D. C., Ceman, S., Nakamoto, M., Mowrey, J., Jongens, T. A., Nelson, D. L., Moses, K. and Warren, S. T. Biochemical and genetic interaction between the fragile X mental retardation protein and the microRNA pathway. Nat. Neurosci. 7, 113-117 (2004).
Joost Haasnoot, P. C., Cupac, D. and Berkhout, B. Inhibition of virus replication by RNA interference. J. Biomed. Sci. 10, 607-616 (2003).
Kamath, R. S., Fraser, A. G., Dong, Y., Poulin, G., Durbin, R., Gotta, M., Kanapin, A., Le Bot, N., Moreno, S., Sohrmann, M., Welchman, D. P., Zipperlen, P. and Ahringer, J. Systematic functional analysis of the Caenorhabditis elegans genome using RNAi. Nature 421, 231-237 (2003).
Kapadia, S. B., Brideau-Andersen, A. and Chisari, F. V. Interference of hepatitis C virus RNA replication by short interfering RNAs. Proc. Natl. Acad. Sci. USA. 100, 2014-2018 (2003).
Karagiannis, T. C. and El-Osta, A. RNA interference and potential therapeutic applications of short interfering RNAs. Cancer Gene Ther. (2005).
Kartasheva, N. N., Contente, A., Lenz-Stoppler, C., Roth, J. and Dobbelstein, M. p53 induces the expression of its antagonist p73 Delta N, establishing an autoregulatory feedback loop. Oncogene 21, 4715-4727 (2002).
Kawasaki, H., Suyama, E., Iyo, M. and Taira, K. siRNAs generated by recombinant human Dicer induce specific and significant but target site-independent gene silencing in human cells. Nucleic Acids Res. 31, 981-987 (2003).
Kennerdell, J. R. and Carthew, R. W. Heritable gene silencing in Drosophila using double-stranded RNA. Nat. Biotechnol. 18, 896-898 (2000).
Khvorova, A., Reynolds, A. and Jayasena, S. D. Functional siRNAs and miRNAs exhibit strand bias. Cell 115, 209-216 (2003).
Kittler, R. and Buchholz, F. RNA interference: gene silencing in the fast lane. Semin. Cancer Biol. 13, 259-265 (2003).
Klein, C., Bock, C. T., Wedemeyer, H., Wustefeld, T., Locarnini, S., Dienes, H. P., Kubicka, S., Manns, M. P. and Trautwein, C. Inhibition of hepatitis B virus replication in vivo by nucleoside analogues and siRNA. Gastroenterology 125, 9-18 (2003).
Kong, X. C., Barzaghi, P. and Ruegg, M. A. Inhibition of synapse assembly in mammalian muscle in vivo by RNA interference. EMBO Rep. 5, 183-188 (2004).
Kretschmer-Kazemi Far, R. and Sczakiel, G. The activity of siRNA in mammalian cells is related to structural target accessibility: a comparison with antisense oligonucleotides. Nucleic Acids Res. 31, 4417-4424 (2003).
Kuiken, T., Fouchier, R., Rimmelzwaan, G. and Osterhaus, A. Emerging viral infections in a rapidly changing world. Curr. Opin. Biotechnol. 14, 641-646 (2003).
Kumar, R., Conklin, D. S. and Mittal, V. High-throughput selection of effective RNAi probes for gene silencing. Genome Res. 13, 2333-2340 (2003).
Lassus, P., Opitz-Araya, X. and Lazebnik, Y. Requirement for caspase-2 in stress-induced apoptosis before mitochondrial permeabilization. Science 297, 1352-1354 (2002).
Lee, N. S., Dohjima, T., Bauer, G., Li, H., Li, M. J., Ehsani, A., Salvaterra, P. and Rossi, J. Expression of small interfering RNAs targeted against HIV-1 rev transcripts in human cells. Nat. Biotech. 20, 500-505 (2002).
Lee, Y., Ahn, C., Han, J., Choi, H., Kim, J., Yim, J., Lee, J., Provost, P., Radmark, O., Kim, S. and Kim, V. N. The nuclear RNase III Drosha initiates microRNA processing. Nature 425, 415-419 (2003).
Lee, Y., Kim, M., Han, J., Yeom, K. H., Lee, S., Baek, S. H. and Kim, V. N. MicroRNA genes are transcribed by RNA polymerase II. EMBO J. 23, 4051-4060 (2004).
Lee, Y. S., Nakahara, K., Pham, J. W., Kim, K., He, Z., Sontheimer, E. J. and Carthew, R. W. Distinct roles for Drosophila Dicer-1 and Dicer-2 in the siRNA/miRNA silencing pathways. Cell 117, 69-81 (2004).
Lewis, D. L., Hagstrom, J. E., Loomis, A. G., Wolff, J. A. and Herweijer, H. Efficient delivery of siRNA for inhibition of gene expression in postnatal mice. Nat. Genet. 32, 107-108 (2002).
Liu, C. M., Liu, D. P., Dong, W. J. and Liang, C. C. Retrovirus vector-mediated stable gene silencing in human cell. Biochem. Biophys. Res. Commun. 313, 716-720 (2004).
Lobo, S. M. and Hernandez, N. A 7 bp mutation converts a human RNA polymerase II snRNA promoter into an RNA polymerase III promoter. Cell 58, 55–67 (1989).
Lobo, S. M. and Hernandez, N. Trancription of snRNA genes by RNA polymerases II and III. Transcription, Mechanisms and Regulation. pp. 127-159 (1994).
Lobo, S. M., Ifill, S. and Hernandez, N. cis-acting elements required for RNA polymerase II and III transcription in the human U2 and U6 snRNA promoters. Nucleic Acids Res. 18, 2891-2899 (1990).
Lu, J., Getz, G., Miska, E. A., Alvarez-Saavedra, E., Lamb, J., Peck, D., Sweet-Cordero, A., Ebert, B. L., Mak, R. H., Ferrando, A. A., Downing, J. R., Jacks, T., Horvitz, H. R. and Golub, T. R. MicroRNA expression profiles classify human cancers. Nature 435, 834-838 (2005).
Lu, P. Y., Xie, F. Y. and Woodle, M. C. siRNA-mediated antitumorigenesis for drug target validation and therapeutics. Curr. Opin. Mol. Ther. 5, 225-234 (2003).
Lu, P. Y., Xie, F. Y. and Woodle, M. C. Modulation of angiogenesis with siRNA inhibitors for novel therapeutics. Trends Mol. Med. 11, 104-113 (2005).
Luciano, D. J., Mirsky, H., Vendetti, N. J. and Maas, S. RNA editing of a miRNA precursor. RNA 10, 1174-1177 (2004).
Lund, E., Guttinger, S., Calado, A., Dahlberg, J. E. and Kutay, U. Nuclear export of microRNA precursors. Science 303, 95-98 (2004).
Marshall, E. Gene therapy. Second child in French trial is found to have leukemia. Science 299, 320 (2003).
Martinez, J., Patkaniowska, A., Urlaub, H., Luhrmann, R. and Tuschl, T. Single-stranded antisense siRNAs guide target RNA cleavage in RNAi. Cell 110, 563–574 (2002).
Matsuda, T. and Cepko, C. L. Electroporation and RNA interference in the rodent retina in vivo and in vitro. Proc. Natl. Acad. Sci. USA. 101, 16-22 (2003).
Matsukura, S., Jones, P. A., and Takai, D. Establishment of conditional vectors for hairpin siRNA knockdowns. Nucleic Acid Res. 31, e77 (2003).
Mattaj, I., Dathan, N. A., Parry, H. D., Carbon, P. and Krol, A. Changing the RNA polymerase specificity of U6 snRNA gene promoters. Cell 55, 435-442 (1988).
McCaffrey, A. P., Meuse, L., Pham, T. T., Conklin, D. S., Hannon, G. J. and Kay, M. A. RNA interference in adult mice. Nature 418, 38-39 (2002).
McCaffrey, A. P., Nakai, H., Pandey, K., Huang, Z., Salazar, F. H., Xu, H., Wieland, S. F., Marion, P. L., Kay, M. A. Inhibition of hepatitis B virus in mice by RNA interference. Nat. Biotechnol. 21, 639-644 (2003).
McDonnell, S., Navre, M., Coffey, R. J., Jr. and Matrisian, L. M. Expression and localization of the matrix metalloproteinase pump-1 (MMP-7) in human gastric and colon carcinomas. Mol. Carcinog. 4, 527-533 (1991).
McManus, M. T., Petersen, C. P., Haines, B. B., Chen, J. and Sharp, P. A. Gene silencing using micro-RNA designed hairpins. RNA 8, 842-850 (2002).
Mellitzer, G., Hallonet, M., Chen, L. and Ang, S. Spatial and temporal ‘knock down’ of gene expression by electroporation of double-stranded RNA and morpholinos into early postimplantation mouse embryos. Mech. Dev. 118, 57 (2002).
Mette, M. F., Aufsatz, W., van der Winden, J., Matzke, M. A. and Matzke, A. J. Transcriptional silencing and promoter methylation triggered by double-stranded RNA. EMBO J. 19, 5194-5201 (2000).
Mertens, C. and Rulicke, T. A comprehensive form for the standardized characterization of transgenic rodents: genotype, phenotype, welfare assessment, recommendations for refinement. ALTEX. 17, 15-21 (2000).
Michael, M. Z., O' Connor, S. M., van Holst Pellekaan, N. G., Young, G. P. and James, R. J. Reduced accumulation of specific microRNAs in colorectal neoplasia. Mol. Cancer Res. 1, 882-891 (2003).
Miyagishi, M. and Taira, K. U6 promoter-driven siRNAs with four uridine 3' overhangs efficiently suppress targeted gene expression in mammalian cells. Nat. Biotechnol. 20, 497-500 (2002).
Morens, D. M., Folkers, G. K. and Fauci, A. S. The challenge of emerging and re-emerging infectious diseases. Nature 430, 242-249 (2004).
Mousses, S., Caplen, N. J., Cornelison, R., Weaver, D., Basik, M., Hautaniemi, S., Elkahloun, A. G., Lotufo, R. A., Choudary, A., Dougherty, E. R., Suh, E. and Kallioniemi, O. RNAi microarray analysis in cultured mammalian cells. Genome Res. 13, 2341-2347 (2003).
Muller, D., Breathnach, R., Engelmann, A., Millon, R., Bronner, G., Flesch, H., Dumont, P., Eber, M. and Abecassis, J. Expression of collagenase-related metalloproteinase genes in human lung or head and neck tumours. Int. J. Cancer 48, 550-556 (1991).
Murphy, S., Di Liegro, C. and Melli, M. The in vitro transcription of the 7SK RNA gene by RNA polymerase III is dependent only on the presence of an upstream promoter. Cell 51, 81-87 (1987).
Murphy, S., Pierani, A., Scheidereit, C., Melli, M. and Roeder, R. G. Purified octamer binding transcription factors stimulate RNA polymerase III--mediated transcription of the 7SK RNA gene. Cell 59, 1071-1080 (1989).
Murphy, S., Yoon, J.-B., Gerster, T. and Roeder, R. G. Oct-1 and Oct-2 potentiate functional interactions of a transcription factor with the proximal sequence element of small nuclear RNA genes. Mol. Cell Biol. 12, 3247-3261 (1992).
Myslinski, E., Ame, J. C., Krol, A. and Carbon, P. An unusually compact external promoter for RNA polymerase  transcription of the human H1RNA gene. Nucleic Acids Res. 29, 2502-2509 (2001).
Myslinski, E., Krol, A. and Carbon, P. Optimal tRNASec gene activity requires an upstream SPH motif. Nucleic Acids Res. 20, 203–209 (1992).
Nagashima, Y., Hasegawa, S., Koshikawa, N., Taki, A., Ichikawa, Y., Kitamura, H., Misugi, K., Kihira, Y., Matuo, Y., Yasumitsu, H. and Miyazaki, K. Expression of matrilysin in vascular endothelial cells adjacent to matrilysin-producing tumors. Int. J. Cancer 72, 441-445 (1997).
Naito, Y., Yamada, T., Ui-Tei, K., Morishita, S. and Saigo, K. siDirect: highly effective, target-specific siRNA design software for mammalian RNA interference. Nucleic Acids Res. 32, W124-129 (2004).
Newell, K. J., Witty, J. P., Rodgers, W. H. and Matrisian, L. M. Expression and localization of matrix-degrading metalloproteinases during colorectal tumorigenesis. Mol. Carcinog. 10, 199-206 (1994).
Novina, C. D., Murray, M. F., Dykxhoorn, D. M., Beresford, P. J., Riess, J., Lee, S. K., Collman, R. G., Lieberman, J., Shankar, P. and Sharp, P. A. siRNA-directed inhibition of HIV-1 infection. Nat. Med. 8, 681-686 (2002).
O'Donnell, K. A., Wentzel, E. A., Zeller, K. I., Dang, C. V. and Mendell, J. T. c-Myc-regulated microRNAs modulate E2F1 expression. Nature 435, 839-843 (2005).
Overhoff, M., Alken, M., Far, R. K., Lemaitre, M., Lebleu, B., Sczakiel, G. and Robbins, I. Local RNA target structure influences siRNA efficacy: a systematic global analysis. J. Mol. Biol. 348, 871-881 (2005).
Paddison, P. J., Caudy, A. A, Bernstein, E., Hannon, G. J. and Conklin, D. S. Short hairpin RNAs (shRNAs) induce sequence-specific silencing in mammalian cells. Genes Dev. 16, 948-958 (2002).
Paddison, P. J., Caudy, A. A. and Hannon, G. J. Stable suppression of gene expression by RNAi in mammalian cells. Proc. Natl. Acad. Sci. U S A. 99, 1443-1448 (2002).
Paddison, P. J. and Hannon, G. J. RNA interference: the new somatic cell genetics? Cancer Cell 2, 17-23 (2002).
Paddison, P. J., Silva, J. M., Conklin, D. S., Schlabach M, Li M, Aruleba S, Balija V, O'Shaughnessy A, Gnoj L, Scobie K, Chang K, Westbrook T, Cleary M, Sachidanandam R, McCombie WR, Elledge SJ, and Hannon, G. J. A resource for large-scale RNA-interference-based screens in mammals. Nature 428, 427-431 (2004).
Pal-Bhadra, M., Bhadra, U. and Birchler, J. A. RNAi relaed mechanisms affect both transcriptional and posttranscriptional transgene silencing in Drosophila. Mol. Cell 9, 315-327 (2002).
Park, W. S., Miyano-Kurosaki, N., Hayafune, M., Nakajima, E., Matsuzaki, T., Shimada, F. and Takaku, H. Prevention of HIV-1 infection in human peripheral blood mononuclear cells by specific RNA interference. Nucleic Acids Res. 30, 4830-4835 (2002).
Parry, H. D., Scherly, D. and Mattaj, I. W.‘Snurpogenesis’: The transcription and assembly of U snRNP components. Trends Biochem. Sci. 14, 15-19 (1989).
Paul, C. P., Good, P. D., Winer, I. and Engelke, D. R. Effective expression of small interfering RNA in human cells. Nat. Biotechnol. 20, 505-508 (2002).
Paule, M. R. and White, R. J. Survey and summary: transcription by RNA polymerases I and III. Nucleic Acids Res. 28, 1283-1298 (2000).
Pothof, J., van Haaften, G., Thijssen, K., Kamath, R. S., Fraser, A. G., Ahringer, J., Plasterk, R. H. and Tijsterman, M. Identification of genes that protect the C. elegans genome against mutations by genome-wide RNAi. Genes Dev. 17, 443-448 (2003).
Qin, X. F., An, D. S., Chen, I. S. and Baltimore, D. Inhibiting HIV-1 infection in human T cells by lentiviral-mediated delivery of small interfering RNA against CCR5. Proc. Natl. Acad. Sci. U S A. 100, 183-188 (2003).
Reich, S. J., Fosnot, J., Kuroki, A., Tang, W., Yang, X., Maguire, A. M., Bennett, J. and Tolentino, M. J. Small interfering RNA (siRNA) targeting VEGF effectively inhibits ocular neovascularization in a mouse model. Mol. Vis. 9, 210-216 (2003).
Reynolds, A., Leake, D., Boese, Q., Scaringe, S., Marshall, W. S. and Khvorova, A. Rational siRNA design for RNA interference. Nat. Biotechnol. 22, 326-330 (2004).
Rubinson, D. A., Dillon, C. P., Kwiatkowski, A. V., Sievers, C., Yang, L., Kopinja, J., Rooney, D. L., Ihrig, M. M, McManus, M. T., Gertler, F. B., Scott, M. L. and Van Parijs, L. A lentivirus-based system to functionally silence genes in primary mammalian cells, stem cells and transgenic mice by RNA interference. Nat. Genet. 33, 401-406 (2003).
Ruiz, M. T., Voinnet, O. and Baulcombe, D. C. Initiation and maintenance of virus-induced gene silencing. Plant Cell 10, 937-946 (1998).
Sadowski, C. L., Henry, R. W., Lobo, S. M. and Hernandez, N. Targeting TBP to a non-TATA box cis-regulatory element: a TBP-containing complex activates transcription from snRNA promoters through the PSE. Genes Dev. 7, 1535-1548 (1993).
Safinya, C. R. Structures of lipid-DNA complexes: supramolecular assembly and gene delivery. Curr. Opin. Struct. Biol. 11, 440-448 (2001).
Saksela, K. Human viruses under attack by small inhibitory RNA. Trends Microbiol. 11, 345-347 (2003).
Schaub, M., Myslinski, E., Schuster, C., Krol, A. and Carbon, P. Staf, a promiscuous activator for enhanced transcription by RNA polymerases II and III. EMBO J. 16, 173-181 (1997).
Scherr, M., Battmer, K., Ganser, A. and Eder, M. Modulation of gene expression by lentiviral-mediated delivery of small interfering RNA. Cell Cycle 2, 251-257 (2003).
Schiffelers, R. M., Ansari, A., Xu, J., Zhou, Q., Tang, Q., Storm, G., Molema, G., Lu, P. Y., Scaria, P. V. and Woodle, M. C. Cancer siRNA therapy by tumor selective delivery with ligand-targeted sterically stabilized nanoparticle. Nucleic Acids Res. 32, e149 (2004).
Schubert, S., Grunweller, A., Erdmann, V. A. and Kurreck, J. Local RNA target structure influences siRNA efficacy: systematic analysis of intentionally designed binding regions. J. Mol. Biol. 348, 883-893 (2005).
Schuster, C., Myslinski, E., Krol, A. and Carbon, P. Staf, a novel zinc finger protein that activates the RNA polymerase III promoter of the selenocysteine tRNA gene. EMBO J. 14, 3777-3787 (1995).
Schwarz, D. S., Hutvagner, G., Du, T., Xu, Z., Aronin, N. and Zamore, P. D. Asymmetry in the assembly of the RNAi enzyme complex. Cell 115, 199-208 (2003).
Schwarz, D. S., Hutvagner, G., Haley, B. and Zamore, P. D. Evidence that siRNAs function as guides, not primers, in the Drosophila and human RNAi pathways. Mol. Cell 10, 537-548 (2002).
Seggerson, K., Tang, L. and Moss, E. G. Two genetic circuits repress the Caenorhabditis elegans heterochronic gene lin-28 after translation initiation. Dev. Biol. 243, 215-225 (2002).
Shen, C., Buck, A. K., Liu, X., Winkler, M. and Reske, S. N. Gene silencing by adenovirus-delivered siRNA. FEBS Lett. 539, 111-114 (2003).
Shigemasa, K., Tanimoto, H., Sakata, K., Nagai, N., Parmley, T. H., Ohama, K. and O’Brien, T. J. Induction of matrix metalloprotease-7 is common in mucinous ovarian tumors including early stage disease. Med. Oncol. 17, 52-58 (2000).
Shlomai, A. and Shaul, Y. Inhibition of hepatitis B virus expression and replication by RNA interference. Hepatology 37, 764-770 (2003).
Shu, X., Wu, W., Mosteller, R. D. and Broek, D. Sphingosine kinase mediates vascular endothelial growth factor-induced activation of ras and mitogen-activated protein kinases. Mol. Cell Biol. 22, 7758-7768 (2002).
Shuey, D. J., McCallus, D. E., and Giordano, T. RNAi：gene-silencing in the therapeutic intervention. DDT. 7, 1040-1046 (2002).
Sijen, T., Fleenor, J., Simmer, F., Thijssen, K. L., Parrish, S., Timmons, L., Plasterk, R. H. and Fire, A. On the role of RNA amplification in dsRNA-triggered gene silencing. Cell 107, 465-76 (2001).
Silva, J., Chang, K., Hannon, G. J. and Rivas, F. V. RNA-interference-based functional genomics in mammalian cells: reverse genetics coming of age. Oncogene 23, 8401-8409 (2004).
Siolas, D., Lerner, C., Burchard, J., Ge, W., Linsley, P. S., Paddison, P. J., Hannon, G. J. and Cleary, M. A. Synthetic shRNAs as potent RNAi triggers. Nat. Biotechnol. 23, 227-231 (2005).
Sioud, M. Therapeutic siRNAs. Trends Pharmacol. Sci. 25, 22-28 (2004).
Smart, N., Scambler, P. J. and Riley, P. R. A rapid and sensitive assay for quantification of siRNA efficiency and specificity. Biol. Proced. Online 7, 1-7 (2005).
Song, E., Lee, S. K., Dykxhoorn, D. M., Novina, C., Zhang, D., Crawford, K., Cerny, J., Sharp, P. A., Lieberman, J., Manjunath, N. and Shankar, P. Sustained small interfering RNA-mediated human immunodeficiency virus type 1 inhibition in primary macrophages. J. Virol. 77, 7174-7181 (2003).
Song, E., Lee, S. K., Wang, J., Ince, N., Ouyang, N., Min, J., Chen, J., Shankar, P. and Lieberman, J. RNA interference targeting Fas protects mice from fulminant hepatitis. Nat. Med. 9, 347-351 (2003).
Song, E., Zhu, P., Lee, S. K., Chowdhury, D., Kussman, S., Dykxhoorn, D. M., Feng, Y., Palliser, D., Weiner, D. B., Shankar, P., Marasco, W. A. and Lieberman, J. Antibody mediated in vivo delivery of small interfering RNAs via cell-surface receptors. Nat. Biotechnol. 23, 709-717 (2005).
Sorensen, D. R., Leirdal, M. and Sioud, M. Gene silencing by systemic delivery of synthetic siRNAs in adult mice. J. Mol. Biol. 327, 761–766 (2003).
Soutschek, J., Akinc, A., Bramlage, B., Charisse, K., Constien, R., Donoghue, M., Elbashir, S., Geick, A., Hadwiger, P., Harborth, J., John, M., Kesavan, V., Lavine, G., Pandey, R. K., Racie, T., Rajeev, K. G., Rohl, I., Toudjarska, I., Wang, G., Wuschko, S., Bumcrot, D., Koteliansky, V., Limmer, S., Manoharan, M. and Vornlocher, H. P. Therapeutic silencing of an endogenous gene by systemic administration of modified siRNAs. Nature 432, 173-178 (2004).
Stark, G. R., Kerr, I. M., Williams, B. R., Silverman, R. H. and Schreiber, R. D. How cells respond to interferons. Annu. Rev. Biochem. 67, 227-264 (1998).
Stewart, S. A., Dykxhoorn, D. M., Palliser, D., Mizuno, H., Yu, E. Y., An, D. S., Sabatini, D. M., Chen, I. S., Hahn, W. C., Sharp, P. A., Weinberg, R. A. and Novina, C. D. Lentivirus-delivered stable gene silencing by RNAi in primary cells. RNA. 9, 493-501 (2003).
Surabhi, R. M. and Gaynor, R. B. RNA interference directed against viral and cellular targets inhibits human immunodeficiency Virus Type 1 replication. J. Virol. 76, 12963-12973 (2002).
Svoboda, P., Stein, P. and Schultz, R. M. RNAi in mouse oocytes and preimplantation embryos: effectiveness of hairpin dsRNA. Biochem. Biophys. Res. Commun. 287, 1099-1104 (2001).
Tabara, H., Grishok, A. and Mello, C. C. RNAi in C. elegans: soaking in the genome sequence. Science 282, 430-431 (1998).
Tabara, H., Sarkissian, M., Kelly, W. G., Fleenor, J., Grishok, A., Timmons, L., Fire, A. and Mello, C. C. The rde-1 gene, RNA interference, and transposon silencing in C. elegans. Cell 99, 123-132 (1999).
Takamizawa, J., Konishi, H., Yanagisawa, K., Tomida, S., Osada, H., Endoh, H., Harano, T., Yatabe, Y., Nagino, M., Nimura, Y., Mitsudomi, T. and Takahashi, T. Reduced expression of the let-7 microRNAs in human lung cancers in association with shortened postoperative survival. Cancer Res. 64, 3753-3756 (2004).
Tang, G., Reinhart, B. J., Bartel, D. P. and Zamore, P. D. A biochemical framework for RNA silencing in plants. Genes Dev. 17, 49-63 (2003).
Tavernarakis, N., Wang, S. L., Dorovkov, M., Ryazanov, A. and Driscoll, M. Heritable and inducible genetic interference by double-stranded RNA encoded by transgenes. Nat. Genet. 24, 180-183 (2000).
Templeton, N. S. Cationic liposome-mediated gene delivery in vivo. Biosci. Rep. 22, 238-295 (2002).
Templeton, N. S. Cationic liposomes as in vivo delivery vehicles. Curr. Med. Chem. 10, 1279-1287 (2003).
Timmons, L., Tabara, H., Mello, C. C. and Fire, A. Z. Inducible systemic RNA silencing in Caenorhabditis elegans. Mol. Biol. Cell 14, 2972-2983 (2003).
Tiscornia, G., Singer, O., Ikawa, M. and Verma, I. M. A general method for gene knockdown in mice by using lentiviral vectors expressing small interfering RNA. Proc. Natl. Acad. Sci. U S A. 100, 1844-1848 (2003).
Thomas, C. E., Ehrhardt, A. and Kay, M. A. Progress and problems with the use of viral vectors for gene therapy. Nat. Rev. Genet. 4, 346-358 (2003).
Topper, J. N. and Clayton, D. A. Characterization of human MRP/Th RNA and its nuclear gene: full length MRP/Th RNA is an active endoribonuclease when assembled as an RNP. Nucleic Acids Res. 18, 793-799 (1990).
Tuschl, T. Functional genomics: RNA sets the standard. Nature 421, 220-221 (2003).
Tuschl, T., Zamore, P. D., Lehmann, R., Bartel, D. P. and Sharp, P. A. Targeted mRNA degradation by double-stranded RNA in vitro. Genes Dev. 13, 3191-3197 (1999).
Ueno, H., Yamashita, K., Azumano, I., Inoue, M. and Okada, Y. Enhanced production and activation of matrix metalloproteinase-7 (matrilysin) in human endometrial carcinomas. Int. J. Cancer 84, 470-477 (1999).
Ui-Tei, K., Naito, Y., Takahashi, F., Haraguchi, T., Ohki-Hamazaki, H., Juni, A., Ueda, R. and Saigo, K. Guidelines for the selection of highly effective siRNA sequences for mammalian and chick RNA interference. Nucleic Acids Res. 32, 936-948 (2004).
Van den Haute, C., Eggermont, K., Nuttin, B., Debyser, Z. and Baekelandt, V. Lentiviral vector-mediated delivery of short hairpin RNA results in persistent knockdown of gene expression in mouse brain. Hum. Gene Ther. 14, 1799-1807 (2003).
Vanhecke, D. and Janitz, M. Functional genomics using high-throughput RNA interference. Drug Discov. Today 10, 205-212 (2005).
Ventura, A., Meissner, A., Dillon, C. P., McManus, M., Sharp, P. A., Van Parijs, L., Jaenisch, R. and Jacks, T. Cre-lox-regulated conditional RNA interference from transgenes. Proc. Natl. Acad. Sci. U S A. 101, 10380-10385 (2004).
Vickers, T. A., Koo, S., Bennett, C. F., Crooke, S. T., Dean, N. M. and Baker, B. F. Efficient reduction of target RNAs by small interfering RNA and RNase H-dependent antisense agents. A comparative analysis. J. Biol. Chem. 278, 7108-7118 (2003).
Wall, N. R. and Shi, Y. Small RNA: can RNA interference be exploited for therapy? Lancet 362,1401-1403 (2003).
Wassenegger, M., Heimes, S., Riedel, L. and Sanger, H. L. RNA-directed de novo methylation of genomic sequences in plants. Cell 76, 567-576 (1994).
Wianny, F. and Zernicka-Goetz, M. Specific interference with gene function by double-stranded RNA in early mouse development. Nat. Cell Biol. 2, 70-75 (2000).
Wielockx B, Libert C, Wilson C. Matrilysin (matrix metalloproteinase-7): a new promising drug target in cancer and inflammation? Cytokine Growth Factor Rev. 15, 111-115 (2004).
Wightman, B., Ha, I. and Ruvkun, G. Posttranscriptional regulation of the heterochronic gene lin-14 by lin-4 mediates temporal pattern formation in C. elegans. Cell 75, 855-862 (1993).
Williams, D. A. and Baum, C. Medicine. Gene therapy--new challenges ahead. Science 302, 400-401 (2003).
Wilson, J. A., Jayasena, S., Khvorova, A., Sabatinos, S., Rodrigue-Gervais, I. G., Arya, S., Sarangi, F., Harris-Brandts, M., Beaulieu, S. and Richardson, C. D. RNA interference blocks gene expression and RNA synthesis from hepatitis C replicons propagated in human liver cells. Proc. Natl. Acad. Sci. USA. 100, 2783-2788 (2003).
Wu, M. T., Wu, R. H., Hung, C. F., Cheng, T. L., Tsai, W. H. and Chang, W. T. Simple and efficient DNA vector-based RNAi systems in mammalian cells. Biochem. Biophys. Res. Commun. 330, 53-59 (2005).
Xia, H., Mao, Q., Paulson, H. L. and Davidson, B. L. siRNA-mediated gene silencing in vitro and in vivo. Nat. Biotechnol. 20, 1006-1010 (2002).
Yamashita, K., Azumano, I., Mai, M. and Okada, Y. Expression and tissue localization of matrix metalloproteinase 7 (matrilysin) in human gastric carcinomas. Implications for vessel invasion and metastasis. Int. J. Cancer 79, 187-194 (1998).
Yamashita, K., Mori, M., Shiraishi, T., Shibuta, K. and Sugimachi, K. Clinical significance of matrix metalloproteinase-7 expression in esophageal carcinoma. Clin. Cancer Res. 6, 1169-1174 (2000).
Yang, M., Li, Y. and Padgett, R. W. MicroRNAs: Small regulators with a big impact. Cytokine Growth Factor Rev. (2005).
Yi, R., Qin, Y., Macara, I. G. and Cullen, B. R. Exportin-5 mediates the nuclear export of pre-microRNAs and short hairpin RNAs. Genes Dev. 17, 3011-3016 (2003).
Yoon, H. S., Chen, X. and Yang, V. W. Kruppel-like factor 4 mediates p53-dependent G1/S cell cycle arrest in response to DNA damage. J. Biol. Chem. 278, 2101-2105 (2003).
Yu, J. Y., DeRuiter, S. L. and Turner, D. L. RNA interference by expression of short-interfering RNAs and hairpin RNAs in mammalian cells. Proc. Natl. Acad. Sci. U S A. 99, 6047-6052 (2002).
Yuan, B., Latek, R., Hossbach, M., Tuschl, T. and Lewitter, F. siRNA Selection Server: an automated siRNA oligonucleotide prediction server. Nucleic Acids Res. 32, W130-134 (2004).
Yuan, Y., and Reddy, R. 5' flanking sequences of human MRP/7-2 RNA gene are required and sufficient for the transcription by RNA polymerase III. Biochim. Biophys. Acta 1089, 33-39 (1991).
Zamore, P. D., Tuschl, T., Sharp, P. A. and Bartel, D. P. RNAi: double-stranded RNA directs the ATP-dependent cleavage of mRNA at 21 to 23 nucleotide intervals. Cell 101, 25-33 (2000).
Zender L, Hutker S, Liedtke C, Tillmann HL, Zender S, Mundt B, Waltemathe M, Gosling T, Flemming P, Malek NP, Trautwein C, Manns MP, Kuhnel F, Kubicka S. Caspase 8 small interfering RNA prevents acute liver failure in mice. Proc. Natl. Acad. Sci. U S A. 100, 7797-7802 (2003).
Zeng, Y., and Cullen, B. R. RNA interference in human cells is restricted to the cytoplasm. RNA 8, 855-860 (2002).
Zeng, Y. and Cullen, B. R. Sequence requirements for microRNA processing and function in human cells. RNA 9, 112-123 (2003).
Zeng, Y., Yi, R. and Cullen, B. R. MicroRNAs and small interfering RNAs can inhibit mRNA expression by similar mechanisms. Proc. Natl. Acad. Sci. U S A. 100, 9779-9784 (2003).
Zhang, L., Yang, N., Mohamed-Hadley, A., Rubin, S. C. and Coukos, G. Vector-based RNAi, a novel tool for isoform-specific knock-down of VEGF and anti-angiogenesis gene therapy of cancer. Biochem. Biophys. Res. Commun. 303, 1169-1178 (2003).
Zheng, L., Liu, J., Batalov, S., Zhou, D., Orth, A., Ding S. and Schultz, P. G. An approach to genomewide screens of expressed small interfering RNAs in mammalian cells. Proc. Natl. Acad. Sci. U S A. 101, 135-140 (2004).