genome wide shrna screen - (Feb/13/2011 )
can someone explain what a genome wide shrna screen is? does it target all the genes in the human genome, or how does it work? thanks
The so-called genome wide siRNA or shRNA screen is not targeting all genes, but a single gene. First a library of shRNAs is created that target all genes in the genome. The library is introduced into cells in a control way so that one cell will only get one shRNA at transduction. Then the transduced cells are selected for certain phenotype such as arrest at G1 cell cycle, the shRNA that has been integrated into the DNA will be recovered from the enriched cells, the its identity will be identified by sequencing. Then you will known the knockdown of which gene cause the observed phenotype.
postdoc thank you so much for a great explanation! what is a pooled genome wide siRNA or shRNA screen? where does the pooling part happen and what is it supposed to do? are they just screens of siRNAs or shRNAs targeting multiple genes? thanks!