Dazl shRNA construct design

Dann C T et al. PNAS 2006;103:11246-11251

©2006 by National Academy of Sciences

Transgene expression and DAZL knockdown

Dann C T et al. PNAS 2006;103:11246-11251

©2006 by National Academy of Sciences

Analysis of Dazl-shRNA phenotype

Dann C T et al. PNAS 2006;103:11246-11251

©2006 by National Academy of Sciences

Reverse genetics in the rat: Advantages to using in vivo RNAi over traditional knock-out methods

• Because of the small size of the sequences encoding shRNAs, these sequences can be cloned in lentiviral vectors

• Production of transgenic founders with lentivirus is easy and efficient

• In vivo RNAi allows to generate animals with different degrees of gene deficiency. An allelic series of hypomorphs is often more informative for modeling human disease phenotypes

• Conditional in vivo RNAi approach is now feasable in the rat using tissue specific promoters (RNA polymerase II promoters) driving the expression of shRNAs (embedded in microRNAs)