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. Author manuscript; available in PMC: 2019 May 1.
Published in final edited form as: Curr Protoc Stem Cell Biol. 2018 Apr 26;45(1):e50. doi: 10.1002/cpsc.50

Figure 3. sgRNA expression plasmid generation.

Figure 3

Following digestion with Bbs1 restriction endonuclease, the sticky ends of PX459 plasmid complementary base pair with the designed sgRNA. The target specific protospacer of the sgRNA is ligated directly to an sgRNA scaffold, which complexes with the constitutively expressed spCas9 under a chicken β-actin promoter.