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. Author manuscript; available in PMC: 2015 Aug 7.
Published in final edited form as: Cell Stem Cell. 2014 Jun 12;15(2):215–226. doi: 10.1016/j.stem.2014.05.018

Figure 4. HDR-mediated genome editing.

Figure 4

(A) Schematic of Cas9/gRNA and ssDNA oligo targeting sites at the GATA6 locus. A C>T substitution (green) was introduced in the ssDNA HDR template, generating a new BsgI restriction site (underlined), resulting in an R456C amino acid substitution in GATA6. (B) Strategy of HDR-mediated genome editing. (C and D) T7EI and RFLP assay in HUES8 (C) and BJ iPSCs (D) cotransfected with GATA6 gRNA (Cr8) and ssDNA HDR repair template. (E and F) RFLP analysis (E) and allelic sequence distribution (F) in clones generated with GATA6 gRNA/ssDNA. wt, wild-type; mut, mutation. “R456C + random mut” includes clones with undesired mutations in addition to the R456C modification in one or both alleles. (G) Representative sequences of one homozygous (R456C/ R456C) and one compound heterozygous (R456C/R456C + additional mutations) GATA6 mutant clone.