Fig. 1

Efficient generation of knockout hPSC lines using CRISPR-Cpf1. a A scheme of the experimental procedure for generating knockout hPSC lines. b Schematic of Cpf1 crRNA targeting sites at ALKBH1 and CLEC16A loci showing exon structures (green boxes), PCR amplicons (light gray boxes), and restriction sites used for PCR analysis. crRNA targeting sequences are in bold; PAM sequences are in red. c T7EI assay for crRNAs of ALKBH1 and CLEC16A in MEL1 hESCs. The Indel frequency was calculated using the expected fragments. d T7EI assay for crRNAs of ALKBH1 in H1 hESCs and hiPSCs. The Indel frequency was calculated using the expected fragments. e PCR analysis upon crRNA transfection. For ALKBH1, two crRNAs were transfected together. Clones with gene knockout in one allele are in blue, and clones with gene knockout in two alleles are in red. More detailed description and explanation of the band pattern can be found in Supplementary Fig. 2. f Sequencing results of the targeted allele in ALKBH1 and CLEC16A knockout hPSC lines. PAM sequences are in red. Restrictive enzyme site is in blue