Table 3 Evaluation of editing accuracy for TaABCC6 and TaNFXL1 using homoeologs with perfect match (A) or mismatched bases (B) to the sgRNA

From: An optimised CRISPR/Cas9 protocol to create targeted mutations in homoeologous genes and an efficient genotyping protocol to identify edited events in wheat

(A)
Gene	On-target gene ID^a	Total mapped reads^b	Reads with modifications	Editing frequency^c
TaABCC6	TraesCS2A01G451300	14,718	1059	7.2
TaABCC6	TraesCS2D01G451100	5337	235	4.4
TaNFXL1 (edited by crCas9)	TraesCS7A01G518800	1780	294	16.50
TaNFXL1 (edited by crCas9)	TraesCS7B01G434700	26,991	4142	15.30
TaNFXL1 (edited by pcoCas9)	TraesCS7A01G518800	3479	357	10.3
TaNFXL1 (edited by pcoCas9)	TraesCS7B01G434700	22,519	1062	4.7

(B)
Gene	Off-target Gene ID^a	Total mapped reads^b		Reads with modifications	Editing frequency^c
Gene	Off-target Gene ID^a	Edited by crCas9	Edited by pcoCas9	Reads with modifications	Editing frequency^c
TaABCC6	TraesCS2B01G472800	15205	N/A	252	1.7
	TraesCS2A01G451500	1722	N/A	0	< 0.06
	TraesCS2D01G451300	1987	N/A	0	< 0.05
TaNFXL1	TraesCS7D01G688900LC	2621	2887	0	< 0.04

N/A not applicable
^aOnly genes for which a specific fragment was PCR-amplified and sequenced are presented here. See Additional file 1 for more details
^bThe number of reads mapping perfectly to the corresponding target or off-target sequence
^cEditing frequency (%) = (reads with modification/(mapped reads + reads with modification)) × 100

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ISSN: 1746-4811

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