Table 2.

Summary of base-editing features for STAT1 mutation generation.

Mutation	Desired trinucleotide change	Single-guide RNA sequencea	Base editor used	Protospacer adjacent motif (PAM) sequence	On-target base-editing (%)	Off-target changes within the editing window (%)	Number of clones screened to obtain the desired mutation in a heterozygous state
E235G (c.704A>G)	GAA>GGA	GATGAACTAGTGGAGTGGAAG	SpCas9 ABEmax	CGG	50	38	39b
K278E (c.832A>G)	AAA>GAA	GCTTAAAAAGTTGGAGGAAT	SpCas9 ABEmax	TGG	67	7	17
P329L (c.986C>T)	CCT>CTT	GCACCCTCAGAGGCCGCTGGT	SpRY CBE4max	CTT	15	10	16
T385M (c.1154C>T)	ACG>ATG	GGGCACGCACACAAAAGTGA	SpG CBE4max	TGA	21	4	14
D517G (c.1550A>G)	GAC>GGC	gTGTGGACCAGCTGAACATGT	SpCas9 ABEmax	TGG	77	2	12
Y701C (c.2102A>G)	TAT>TGT	gTGGATATATCAAGACTGAGT	SpRY ABEmax	TGA	36	15	24

^aNon-template “g” appended to single-guide RNA to facilitate transcription downstream of the U6 promoter designated in lowercase.

^bResultant clone contains a silent bystander nucleotide change within the editing window.