TABLE 3.

Sanger-sequencing of amplicons containing predicted off-target mutations identified by WGS.

Gene	Sample	Sequence (5′-3′)
NFASC	WT	TCA​GCT​GCC​CAG​TGT​GACTTGGG
	4-4	TCA​GCT​GCC​CAG​ $\text{G}$ GT​GAC​TTGGG
	4-5	TCA​GCT​GCC​CAG​ $\text{G}$ GT​GAC​TTGGG
	5-3	TCA​GCT​GCC​CAG​TGT​GAC​TTGGG
	5-4	TCA​GCT​GCC​CAG​ $\text{G}$ GT​GAC​TTGGG
	5-7	TCA​GCT​GCC​CAG​ $\text{G}$ GT​GAC​TTGGG
	5-8	TCA​GCT​GCC​CAG​ $\text{G}$ GT​GAC​TTGGG
	5-9	TCA​GCT​GCC​CAG​ $\text{G}$ GT​GAC​TTGGG
SFMBT2	WT	GCT​GTG​TTT​GCC​TCT​CTCCCAGG
	4-3	GCT​GGC​TTT​GCC​TCT​C–––– $\mathbf{G}$ GG
	4-6	G $\text{N}$ TGG $\text{N}$ TTTGCCTCTCT $\text{N}$ CCAGG
	5-1	$\text{NNN}$ G $\text{NN}$ TT $\text{NN}$ C $\text{NNNNNCN}$ CCAGG
	5-3	GCT​G $\text{GC}$ ​TTT​G––––––––CCAGG
	5-4	$\text{NNG}$ G $\text{CN}$ TT $\text{NN}$ C $\text{A}$ T $\text{T}$ T $\text{T}$ TCCCAGG
	5-5	GCT​G $\text{GC}$ ​TTT​G––––––––CCAGG
	5-6	$\text{NNN}$ G $\text{GC}$ TTT $\text{N}$ CCT $\text{T}$ T $\text{T}$ TCCCAGG
	5-7	GCT​G $\text{GC}$ ​TTT​GCC–––––––C $\mathbf{G}$ GG
	5-8	G $\text{NN}$ G $\text{GA}$ TTTGCC–––––––C $\mathbf{G}$ GG
LIPC	WT	GCT​GTG​CTT​GCC​TCT​TTCCCTGG
	4-5	GCTGTG $–––$ G $–––––$ $\text{N}$ T $\text{NN}$ CTGG
	4-6	G $\text{NN}$ G $\text{N}$ GC $\text{NN}$ GC $\text{N}$ TCTTTCCCTGG
	5-1	GCT​GTG​CTT​GCC​TCT​TTC​CCTGG

WT, wild-type sequence; underline text: predicted cut site; bolded font: PAM, sequence; red font: denotes mutation; dash(-): deletion; N, not conclusive base determination.