Table 4:

Regular PCR primers for predicted off-target site analysis

gRNA	Target	CFD*	Forward (5’ - 3’)	Reverse (5’ - 3’)
CRISPR887200_SGM (TFAP2A)	intergenic: CCDC141-SESTD1	0.53	GTACTGGGTCCTTCCCTTCA	AAGAGTGGGGCAGACTTTGT
	intergenic:RP11–20A20.2-AL157830.1	0.35	AAGTTAGCCTGGGCTTGTGT	GAAGCATCAAGGTCAGTTGTG
	exon:LTN1	0.32	ATCCATGTTCCCAGAGCTTC	GCCCACGCTGATTAAAAGAT
	intergenic: CHP2-PRKCB	0.31	AAAAACAGGGCTGAGAATGG	TCATAGCTCACCGCTCAAAC
	intergenic: AC092017.1-RCOR3	0.31	CACATCCCCAAAGACATGAG	GCTGACATTTCTTGGCTTGA
CRISPR887208_SGM (TFAP2A)	intergenic: ATP6V1G1P7-RPL7P45	0.75	TTCATCTACCTTTGCAGGTTGT	TCCATAGCAGAGGGGAGACT
	intron:METAP1D	0.38	GGTTAGGGCGTTGCCTATAA	GACAGCCATACTGCTTGTTGA
	intron:FAM160A1	0.31	TTTCCGTTTGTAGCAGTTGG	GCATCCTCTCTCAGCACTCA
	intergenic:RP11–91K8.2-SNORA33	0.29	TTCCACCTGCACACATTTTT	TTTCATTTGACAGGCAGAGC
	intergenic: GPRIN3-RP11–115D19.1	0.27	CTTCACCCAGTTTCCCCTAA	ACGCAAACCAAGAATGATGA
CRISPR980378_SGM (AHR)	intergenic: CTC-419K13.1-ENC1	0.29	GAGGCCACAAAACCATACAA	GGACTTGGAGAAAGCCAGAG
	intergenic: ACA64-SNX29	0.27	TGAAGGAAATGAACCAGTGC	GCCACAGCCATTTGCTTAT
	exon: AHRR/PDCD6	0.24	CACCTGACCCAGACCATCT	CAGGACAGAAAGCTTGTCCA
	intron:HECW2	0.21	GGGGGATGAAAAGCATTAAA	TTCTCTGAGTGGTGCTCAGG
	intron:AOX1	0.17	TACACCTGCCGACCAAATAA	TCAATTCTCTGCCCATCAGA

^*Cutting frequency determination (CFD) indicates likeliness of off-target cleavage at this particular site, based on Doench et. al. 2016⁸³