Abstract
RNA interference (RNAi) efficiently induces sequence-specific gene silencing in mammalian cells through short interfering RNA (siRNA) of 21–23 nucleotides synthesized in vitro or expressed by DNA-based vector. However, introduction of siRNA into mammalian cells by transfection limits the application of RNAi, especially when it is necessary to generate long-term gene silencing in vivo. Virus vector-mediated RNAi provides an alternative to transfection. In the present study, we investigated such transduction system and showed that retrovirus vector-mediated RNAi can substantially down-regulate expression of mouse adult β-globin gene in MEL cells. The results suggest that retrovirus vector-delivered RNAi may find its use in functional genomics and in gene therapy.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
Fire, A., Xu, S., Montgomery, M. K., Kostas, S. A., Driver, S. E., and Mello, C. C. (1998) Potent and specific genetic interference by double-stranded RNA in Caenorhabditis elegans. Nature 391, 806–811.
Hannon, G. J. (2002) RNA interference. Nature 418, 244–251.
Denli, A. M. and Hannon, G. J. (2003) RNAi: an evergrowing puzzle. Trends. Biochem. Sci. 28, 196–201.
Shi, Y. (2003) Mammalian RNAi for masses. Trends. Genet. 19, 9–12.
Zamore, P. D. (2001) RNA interference: listening to the sound of silence. Nat. Struct. Biol. 8, 746–750.
Sui, G., Soohoo, C., Affar, E. B., Gay, F., Shi, Y., Forrester, W. C., and Shi, Y. (2002) A DNA vector-based RNAi technology to suppress gene expression in mammalian cells. Proc. Natl. Acad. Sci. USA 99, 5515–5520.
Yokota, T., Sakamoto, N., Enomoto, N., Tanabe, Y., Miyagishi, M., Maekawa, S., Yi L., Kurosaki, M., Taira, K., Watanabe, M., and Mizusawa, H. (2003) Inhibition of intracellular hepatitis C virus replication by synthetic and vector-derived small interfering RNAs. EMBO Rep. 4, 602–608.
Zhang, L., Yang, N., Mohamed-Hadley, A., Rubin, S. C., and Coukos, G. (2003) 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.
Yu, J. Y., DeRuiter, S. L., and Turner D. L. (2002) RNA interference by expression of short-interfering RNAs and hairpin RNAs in mammalian cells. Proc. Natl. Acad. Sci. U S A 99, 6047–6052.
Tilly, G., Chapuis, J., Vilette, D., Laude, H., and Vilotte J. L. (2003) Efficient and specific down-regulation of prion protein expression by RNAi. Biochem. Biophys. Res. Commun. 305, 548–551.
Rubinson, D. A., Dillon, C. P., Kwiatkowski, A. V., Sievers, C., Yang, L., Kopinja, J., Zhang, M., McManus, M. T., Gertler, F. B., Scott, M. L., and Parijs, L. V. (2003) A lentivirus-based system to functionally silence genes in primary mammalian cells, stem cells, transgenic mice by RNA interference. Nat. Genet. 33, 401–406.
Stewart, S. A., Dykxhoom, D. M., Palliser, D., Mizuno, H., Hu, E. Y., An, D. S., Sabatini, D. M., Chen, I. S., Hahn, W. C., Sharp, P. A., Weinberg, R. A., and Novina, C. D. (2003) Lentivirus-delivered stable gene silencing by RNAi in primary cells. RNA 9, 493–501.
Wiznerowicz, M. and Trono, D. (2003) Conditional suppression of cellular genes: lentivirus vector-mediated drug-inducible RNA interference. J. Virol. 77, 8957–8961.
Barton, G. M. and Medzhitov, R. (2002) Retroviral delivery of small interfering RNA into primary cells. Proc. Natl. Acad. Sci. USA 99, 14943–14945.
Dong, W. J., Li, B., Liu, D. P., Zn, ZX., Li, J., Hao, D. L., Guo, ZC., and Liang, C. C. (2003) Evaluation of optimal expression cassette in retrovirus vector for beta-thalassemia gene therapy. Mol. Biotechnol. 24, 127–140.
Dong, W. J., Liu, D. P., Li, J., Guo, ZC., and Liang C. C. (2000) Comparison of two kinds of methods to determine the titer of recombinant retrovirus containing beta-globin gene based on G418 selection. Mol. Biotechnol. 15, 77–81.
Feng, D. X., Liu, D. P., Huang, Y., Wu, L., Li, T. C., Wu, M., Tang, X. B., and Liang, C. C. (2001) The expression of human a-like globin genes in transgenic mice mediated by bacterial artificial chromosome. Proc. Natl. Acad. Sci. USA 98, 15073–15077.
Liu, C. M., Liu, D. P., Dong, W. J., and Liang, C. C. (2004) Retrovirus vector-mediated stable gene silencing in human cell. Biochem. Biophys. Res. Commun. 313: 716–720.
Kunath, T., Gish, G., Lickert, Jones, H. N., Pawson, T., and Rossant, J. (2003) Transgenic RNA interference in ES cell-derived embryos recapitulates a genetic null phenotype. Nat. Biotechnol. 21, 559–561.
Hasuwa, H., Kaseda, K., Einarsdottir, T., and Okabe, M. (2002) Small interfering RNA and gene silencing in transgenic mice and rats. FEBS Lett. 532, 227–230.
Park, W. S., Miyano-Kurosaki, N., Hayafune, M., Nakajima, E., Matsuzaki, T., Shimada, F., and Takaku, H. (2002) Prevention of HIV-1 infection in human peripheral blood mononuclear cells by specific RNA interference. Nucleic Acids Res. 30, 4830–4835.
Brummelkamp, T. R., Bernards, R., and Agami, R. (2002) Stable suppression of tumorigenicity by virus-mediated RNA interference. Cancer Cell. 2, 243–247.
Kapadia, S. B., Brideau-Andersen, A., and Chisari, F. V. (2003) Interference of hepatitis C virus RNA replication by short interfering RNAs. Proc. Natl. Acad. Sci. USA 100, 2014–2018.
McCaffrey, A. P., Nakai, H., Pandey, K., Huang, Z. Salazar, F. H., Xu, H., Wieland, S. F., Marion, P. L., and Kay, M. A. (2003) Inhibition of hepatitis B virus in mice by RNA interference. Nat. Biotechnol. 21, 639–644.
Author information
Authors and Affiliations
Corresponding author
Rights and permissions
About this article
Cite this article
Zhao, N., Zu, ZX., Liu, CM. et al. Knockdown of mouse adult β-globin gene expression in MEL cells by retrovirus vector-mediated RNA interference. Mol Biotechnol 28, 195–199 (2004). https://doi.org/10.1385/MB:28:3:195
Issue Date:
DOI: https://doi.org/10.1385/MB:28:3:195