Figure 3.
Chemical modifications of sgRNA and its application in human cells. (a) Illustration of the conventional 5′ and 3′ end modification (5′&3′-sgRNA) and our new e-sgRNA design. (b) Co-delivery of Cas9 mRNA and sgRNAs targeting GFP into HEK293 cells expressing GFP. FACS analysis was done to determine the number of GFP− cells. (c) Co-delivery of Cas9 mRNA and sgRNA targeting HBB into HEK293 cells. NHEJ events were determined by deep sequencing analysis. (d) The editing frequencies of three top off-target sites of HBB sgRNA were determined by deep sequencing of amplicon. *P < 0.05. #P < 0.05 compared to the 5′&3′ sgRNA treated group (n = 3), error bars as s.d.