Hi All,

This seems to be a fairly common yet widely ignored question to me. Ideally a scrambled shRNA should have the same sequence but in random order, but how widely is it endorsed and how practical is it. I am getting shRNA targeting my gene of interest from sigma, but they only have luciferase scrambled shRNA as the best negative control. I can also see Addgene also has plenty of scrambled shRNAs- can I use any of those as long as the base vector matches or do I need to use one of those tools to generate the random sequence of my targeting shRNA and then filter it for off-targets and finally synthesize it (I know this is the ideal negative control- but is this actually necessary to publish in a decent journal). I would really appreciate advice from experienced PIs/reviewers or someone who got any related comments from reviewers. Thanks a lot.