DOI: 10.1073/pnas.152327299. Short RNA duplexes produced by hydrolysis with <i>Escherichia coli</i> RNase III mediate effective RNA interference in mammalian cells

Short RNA duplexes produced by hydrolysis with Escherichia coli RNase III mediate effective RNA interference in mammalian cells

10.1073/pnas.152327299

Crossref journal-article

Proceedings of the National Academy of Sciences

Proceedings of the National Academy of Sciences (341)

Abstract

Small interfering RNA (siRNA) has become a powerful tool for selectively silencing gene expression in cultured mammalian cells. Because different siRNAs of the same gene have variable silencing capacities, RNA interference with synthetic siRNA is inefficient and cost intensive, especially for functional genomic studies. Here we report the use of Escherichia coli RNase III to cleave double-stranded RNA (dsRNA) into endoribonuclease-prepared siRNA (esiRNA) that can target multiple sites within an mRNA. esiRNA recapitulates the potent and specific inhibition by long dsRNA in Drosophila S2 cells. In contrast to long dsRNA, esiRNA mediates effective RNA interference without apparent nonspecific effect in cultured mammalian cells. We found that sequence-specific interference by esiRNA and the nonspecific IFN response activated by long dsRNA are independent pathways in mammalian cells. esiRNA works by eliciting the destruction of its cognate mRNA. Because of its simplicity and potency, this approach is useful for analysis of mammalian gene functions.

Bibliography

Yang, D., Buchholz, F., Huang, Z., Goga, A., Chen, C.-Y., Brodsky, F. M., & Bishop, J. M. (2002). Short RNA duplexes produced by hydrolysis with Escherichia coli RNase III mediate effective RNA interference in mammalian cells. Proceedings of the National Academy of Sciences, 99(15), 9942â9947.

Authors 7

Dun Yang (first)
Frank Buchholz (additional)
Zhongdong Huang (additional)
Andrei Goga (additional)
Chih-Ying Chen (additional)
Frances M. Brodsky (additional)
J. Michael Bishop (additional)

References 25 Referenced 246

Dates

Type	When
Created	23 years, 1 month ago (July 28, 2002, 6:26 p.m.)
Deposited	3 years, 4 months ago (April 13, 2022, 1:33 a.m.)
Indexed	3 days, 16 hours ago (Aug. 29, 2025, 6:13 a.m.)
Issued	23 years, 1 month ago (July 2, 2002)
Published	23 years, 1 month ago (July 2, 2002)
Published Online	23 years, 1 month ago (July 2, 2002)
Published Print	23 years, 1 month ago (July 23, 2002)

Funders 0

None

BibTeX

@article{Yang_2002, title={Short RNA duplexes produced by hydrolysis with Escherichia coli RNase III mediate effective RNA interference in mammalian cells}, volume={99}, ISSN={1091-6490}, url={http://dx.doi.org/10.1073/pnas.152327299}, DOI={10.1073/pnas.152327299}, number={15}, journal={Proceedings of the National Academy of Sciences}, publisher={Proceedings of the National Academy of Sciences}, author={Yang, Dun and Buchholz, Frank and Huang, Zhongdong and Goga, Andrei and Chen, Chih-Ying and Brodsky, Frances M. and Bishop, J. Michael}, year={2002}, month=jul, pages={9942–9947} }

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