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Published online on January 27, 2003, 10.1073/pnas.0437912100
PNAS | February 18, 2003 | vol. 100 | no. 4 | 1844-1848


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Genetics
 A general method for gene knockdown in mice by using lentiviral vectors expressing small interfering RNA

(gene silencing|transgenesis|functional genomics)

Gustavo Tiscornia*, Oded Singer*, Masahito Ikawadagger , and Inder M. VermaDagger

Laboratory of Genetics, The Salk Institute, 10010 North Torrey Pines Road, La Jolla, CA 92037

Contributed by Inder M. Verma, December 26, 2002

We describe the use of lentiviral vectors expressing small interfering RNAs (siRNAs) to knock down the expression of specific genes in vitro and in vivo. A lentiviral vector capable of generating siRNA specific for GFP after transduction of 293T-GFP cell lines showed no GFP fluorescence. Furthermore, no GFP-specific RNA could be detected. When eggs from GFP-positive transgenic mice were transduced with lentivirus-expressing siGFP virus, reduced fluorescence could be seen in blastocysts. More interestingly, pups from F1 progeny, which expressed siGFP, showed considerably diminished fluorescence and decreased GFP. We propose that an approach of combining transgenesis by lentiviral vectors expressing siRNAs can be used successfully to generate a large number of mice in which the expression of a specific gene(s) can be down-regulated substantially. We believe that this approach of generating "knockdown" mice will aid in functional genomics.

* G.T. and O.S. contributed equally to this work.

dagger Present address: Genome Information Research Center, Osaka University, 3-1, Yamada-Oka, Suita, Osaka 565-0871, Japan.

Dagger To whom correspondence should be addressed. E-mail: verma{at}salk.edu.

www.pnas.org/cgi/doi/10.1073/pnas.0437912100
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