TABLE 1.

Oligonucleotides

Use	ID	Type	Sequence (5′→3′)
Genome editing	G1	Guide RNA	GTTATCTCATTCTTACACAAAGG
	G2	Guide RNA	ATTCTTACACAAAGGACACTGGG
	O1	ssODN	ACTCCTTAACCAGAAGAAAGGACCTTCCCAGTGTCCTT[A]GTGTAAGAATGAGATAACCAAAAGGTAAATAACATGTGTAA
Genotyping	F1a	Forward primer	TGCAGGTAAGTGTAATTTTCATAGG
	F1b	Forward primer	GGACCTTCCCAGTGTCCTT[A]
	R1	Reverse primer	CCGATGTGCATGGTACTGTC
qPCR	F2	Forward primer	TAACCAGAAGAAAGGAC
	R2	Reverse primer	CTACACATCAATTTCTACTT
	P1	Probe, FAM	CAC[a]AAGGAC
	P2	Probe, HEX	C[T]aAGGACACT
	S1	Standard	CCACACAGTGCGACCACATATTTTGCAAATTTTGTATGCTGAAACTCCTTAACCAGAAGAAAGGACCTTCCCAGTGTCCTT[T]GTGTAAGAATGAGATAACCAAAAGGAGCCTACAAGGAAGTGCAAGGTTTAGTCAACTTGTTGAAGAGCTGCTGAAAAT
	S2	Standard	CCACACAGTGCGACCACATATTTTGCAAATTTTGTATGCTGAAACTCCTTAACCAGAAGAAAGGACCTTCCCAGTGTCCTT[A]GTGTAAGAATGAGATAACCAAAAGGAGCCTACAAGGAAGTGCAAGGTTTAGTCAACTTGTTGAAGAGCTGCTGAAAAT
Sequencing	F3	Forward primer	CACTGCATAGGGAAACTGGC
	R3	Reverse primer	GGGACTGATCTAGGGGTGAC
	F4	Forward primer	AACCCTACCCAGAAAGCTCC
	R4	Reverse primer	ACCTGCAGCTGTCTTGAGAT
	F5	Forward primer	GACCCCTTCATTGTCCTCCA
	R5	Reverse primer	TCCCCTAGCTCCCTCATGAT
	F6	Forward primer	AGTCAGTCCACCATGTCAGT
	R6	Reverse primer	GCAGAGCAGACCCATCGATA
	F7	Forward primer	GCACACCACATCTCTCCTCT
	R7	Reverse primer	GCCAAACAACATGCATGACA
	F8	Forward primer	GTTTCTGGTAAGCAAGCCCC
	R8	Reverse primer	GATCAACTTCTGGGCCATGC
	F9	Forward primer	TGCCCCTTCCCATTATCACA
	R9	Reverse primer	ATCCATGGCAAAGGGAGACA
	F10	Forward primer	CCTGTAACTCCAGCTCCCAA
	R10	Reverse primer	GAAGGCAGGAGGTGAAGTCT
	F11	Forward primer	GGCAGCCTCGTTACACAATC
	R11	Reverse primer	GTGCCCCTCAGACTCTTCAT

Note: Nucleotides in bracket, location of introduced T‐to‐A mutation. Allele‐specific quantitative PCR (qPCR) was carried out by using the cycleave PCR technique. This technique uses a chimeric DNA–RNA–DNA probe labeled with a fluorescent dye and a quencher at its ends; if the probe generates a perfect hybrid with the PCR product, it is digested with RNaseH at the RNA–DNA heteroduplex, leading to separation of the quencher and dye and increased fluorescence; any mismatch at or near the heteroduplex does not lead to RNaseH digestion. Lowercase letter, ribonucleotide; uppercase letter, deoxyribonucleotide. FAM, 6‐carboxyfluorescein‐labeled; HEX, hexachlorofluorescein‐labeled; ssODN, single‐stranded oligodeoxynucleotide.