Efficient mutagenesis and genotyping of maize inbreds using biolistics, multiplex CRISPR/Cas9 editing, and Indel-Selective PCR

Table 2 CRISPR/Cas9 editing outcomes for maize NRT1.1 family genes

gRNA target T0 callus event	NRT1.1B1 mutant¹, %²	NRT1.1B2 mutant¹, %²	NRT1.1B3 mutant¹, %²	NRT1.1A2 mutant¹, %²	NRT1.1C2 mutant¹, %²
1	WT	WT³	WT	+1 (C), 46.7	-18, 51.1
1	WT	WT	WT	WT	+2, 45.3
21	WT	WT	WT	+1 (A), 46.5	+1 (C), 54.8
22	CE	WT	WT	+1 (A), 50	+1 (A), 50
22	WT	WT	WT	+1 (T), 50	+1 (T), 50
70	WT	WT	WT	WT	WT
131	WT	CE	WT	+2, 16.2	+1 (T), 21.9
171	WT	WT	WT	+1 (A), 46.7	+1 (T), 8.4
229	WT	WT	WT	-5, 45.1	WT
233	-3, 100	+1 (A), 100	+1 (T), 100	+1 (C), 100	+1 (T), 100

¹ Sequence change, WT = wild-type sequence, CE = chimeric edit
² Proportion of edited DNA estimated by TIDE analysis
Bold font indicates mutation was also detected in Sanger sequencing of PCR amplicons from at least two of three regenerated T0 plants
³TIDE analysis of callus did not detect edit, but a + 1 (A) mutation was detected in two of three regenerated T0 plants

ISSN: 1746-4811