Fig. 1: CRISPR gene editing correction of the APC germline mutation within FAP-hESC lines.

A CRISPR gene editing correction of the APC germline mutation within FAP1-hESCs. a Strategy to correct the FAP1 mutation. Blue asterisks indicate silent mutations; FAP1 point mutation is marked with red. b Confirmation of APC mutation correction using RFLP. Corrected clones are expected to result in 89 bp and 60 bp for FAP1 and 153 bp for FAP2. Control (H9 line) and FAP(1 + 2) hESC lines served as positive and negative controls, respectively; cut with restriction enzyme: NurI (for FAP1) and BsaJI (for FAP2). c Sequencing results of the APC mutation region following CRISPR editing. The Hues13 hESC line served as a control for non-mutated APC; FAP1 for the heterozygous mutated APC and clone 37 for the corrected clone. B CRISPR gene editing correction of the APC germline mutation within FAP2-hESCs. a FAP2 mutation ata>gta is marked with red; silent mutations are marked in blue; protospacer adjacent motif (PAM) is underlined in purple. b Restriction fragment length polymorphism (RFLP) analysis of PCR products for APC mutation correction. Corrected clones are expected to result in 89 bp and 60 bp for FAP1 and 153 bp for FAP2. Control (H9 line) and FAP1 & FAP2 hESC lines served as controls. The restriction enzyme (E) NurI was used for FAP1, and BsaJI for FAP2. c Sequencing results of the APC mutation region following CRISPR editing. The H9 hESC line served as a control for non-mutated APC; FAP2 for the heterozygous mutated APC.