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Using RNAi
to identify new genes required
for
RNAi  RNA interference (RNAi) is a phenomenon in
which double-stranded RNA (dsRNA) silences endogenous gene expression
in a sequence-specific manner. RNAi shares a remarkable degree of
similarity with silencing phenomena in other organisms, suggesting that
RNAi is part of an ancient and conserved pathway used to regulate gene
expression, to eliminate defective RNAs and to defend against viral
infections and transposons. RNAi has also been implicated in
developmental processes, suggesting that RNAi may play a broader role
in regulating normal gene expression. Since its discovery, the
use of RNAi has become widely employed in many organisms to
specifically knock down gene function. Although we have learned
much about the general mechanisms underlying RNAi, we do not yet have a
complete understanding of how RNAi works.I developed a candidate screen approach using RNAi to identify genes required for RNAi. I identified a number of potential candidates and in a secondary screen, using mutant animals, validated roles for several new genes in RNAi. Interestingly, many of these genes encode protein known to function in the nucleus. In addition, I further characterized the role of a nuclear RNA helicase that functions in RNAi. These results have suggested clues and provided important tools toward furthering our understanding of nuclear mechanisms of RNAi interference in animals.
Protocols: dsRNA preparation (pdf file) back |