Question

I was assigned the gene LOC_Os05g38000

I was asked to design guide RNAs that would remove the whole gene (but make sure primers would still bind). Then draw an image with guide locations and show the size of the deletion. Justify why would you chose those two guides.

Someone please help I'll upvote! Anything will be apreciated!!

Answer 1

Guide RNA used in Crisper/Cas9 technology to modify the gene of interest precisely. Guide RNA contains two different sequences: targeting sequence and Cas9 nuclease-recruiting sequence. Targeting sequence binds with a specific sequence in the gene of interest. Cas9 nuclease-recruiting sequence binds with a nuclease. Upon binding of guide RNA to the gene of interest, Cas9 nuclease cleaves off the DNA molecule. For every gene, we need to design at least four guides RNA and we need to empirically decide which one works will better.