Abstract
To overcome embryonic lethality in disruption of gene function, we developed tissue-specific RNAi vectors and tested under various dsRNA sequences and differential splicing efficiency of intron-spacer. We used Wx locus as a target gene to test various RNAi vectors. At first, pWRI-A was constructed from endosperm-specific Wx promoter, the 5'UTR sense and antisense Wx sequences with intron-spacer and led to complete suppression of Wx expression in endosperm. Further studies examined and results showed that the efficient splicing led to the most effective suppression of target gene expression. dsRNA homologous to coding region trigger complete Wx suppression in both endosperm and pollen, whereas the 5'UTR dsRNA trigger suppression only in endosperm. Based on these results, we are developing various cassette vectors for tissue-specific RNAi to apply for functional analysis of gene shows pleiotropic expression or lethality caused by gene disruption.
