I am using shRNAs to knockdown target genes. But I have been having trouble observing the knockdown. I have 2 projects going on. shRNAs are validated by Luciferase Assay. In one project, the selected shRNA failed to show any reduction in protein expression levels in WB. I then infected the cell line with 5 different shRNA constructs and still none of them showed any reduction. Of the 5, 3 showed good efficiency in luciferase assay. In the other project, we selected an shRNA from the assay and generated a mouse using lentiviral transgenesis. The construct has the marker protein and the shRNA under separate promoters. The mouse expresses the marker protein in almost all cells abut can still produce loads of the target protein. I am really confused and would appreciate some help. Could anyone please give me some advice? Thanks
Edited by zodiac1505, 19 August 2009 - 01:32 AM.