Generation of SVEP1-/- iPSC lines
An isogenic pair of SVEP1 GM23720 iPSC line was generated by
CRISPR genome editing in collaboration with Horizon Discovery Ltd. A
guide RNA targetting
GAGACCGCGCCCGGGGCCCCCGGGAGTATCCCCGCGCCG CCCGCTCCTGGCGA, a region
within exon 1 of Ensembl SVEP1 transcript SVEP1-003 (ENST00000374469.5)
was designed. The underlined highlighted sequence indicates the
protospacer adjacent motif and the italic sequence indicates the guide
RNA. This guide RNA was co-transfected into iPSCs with a plasmid
expressing CAS9. After transfection, iPSCs were serially diluted into 96
well plates to generate single cell clones. Single cell clones were
genotyped by sequencing PCR products generated using primers
CAGCCGCTCTGTCTCCAG and AGGAGATGGCAGGGATCTCT.