Figure 4.

Knock-in of green fluorescent protein (GFP) expression cassette into pp38 of HP8-Cas9. (a) Schematics of GFP expression cassette and pp38 with the gRNA targeting sites at N and C terminus. (b) Schematics of the anticipated pp38 with inserted GFP expression cassette. GFP could be inserted in either orientation. As a result, there are two potential products for each target site. The location of primers used to confirm the presence and the orientation of GFP insert by junction PCR are shown. (c) Junction PCR products with the expected size of the mixed GFP positive cells using the primers shown in Figure 4b. GFP was successfully inserted into pp38 at both sites with both orientations.