Fig. 2.

CRISPR/Cas9-mediated insertion of LDLR expression cassette at the AAVS1 genomic site. a Design of the LDLR expression cassette in the AAVS1 SA-2A-puro-pA donor plasmid that contains sequences homologous to the sequences flanking the AAVS1 double strand break (DSB) induced by CRISPR/Cas9 (5′ and 3′ Arms). Abbreviations: AAVS1, adeno-associated virus integration site 1, APOA2, apolipoprotein A2; pA, polyadenylation tail; Puro(R), puromycin resistance gene; SA, splice acceptor site; WPRE, woodchuck posttranslational regulatory element. b Junction PCR analyses showing correct integration of the cassette in 15/16 FH-iPSC clones. c PCR analysis corresponding to AAVS1 targeted locus, showing absence of integration of the correcting cassette in one of the alleles