Efficient gene editing in Corynebacterium glutamicum using the CRISPR/Cas9 system

Table 2 Results of the porB deletion in C. glutamicum ATCC 13032 and C. glutamicum CGMCC1.15647

No	Host cell	Plasmid	Element	Results (D/W/T)	Efficiency (%)
1	ATCC 13032	pFSC	cas9	0/5/5	0
2	ATCC 13032	pFST-porBsgRNA	sgRNA	0/5/5	0
3	ATCC 13032	pFST-porBT	Hdarm	0/5/5	0
4	ATCC 13032	pFSC, pFST-porBT	Cas9 + Hdarm	0/5/5	0
5	ATCC 13032	pFSC, pFST-porBsgRNA	Cas9 + sgRNA	0/0/0	0
6	ATCC 13032	pFST-porB	sgRNA + Hdarm	0/6/6	0
7	ATCC 13032	pFSC, pFST-porB	Cas9 + sgRNA + Hdarm (1000 bp)	18/0/18	100
8	CGMCC 1.15647	pFSC, pFST-porB15647	Cas9 + sgRNA + Hdarm (1000 bp)	16/0/16	100
9	ATCC 13032	pFSC, pFST-porB600	Cas9 + sgRNA + Hdarm (600 bp)	10/2/12	83.3
10	ATCC 13032	pFSC, pFST-porB300	Cas9 + sgRNA + Hdarm (300 bp)	10/2/12	83.3
11	ATCC 13032	pFSC, pFST-porB100	Cas9 + sgRNA + Hdarm (100 bp)	2/10/12	16.7

D, number of colonies that harbored gene-deleted cells; W, number of colonies that harbored wild type cells; T, total number of colonies used for PCR screening; Efficiency, probability of deletion events occurring, calculated as D/T * 100%; ATCC 13032, C. glutamicum ATCC 13032; CGMCC 1.15647, C. glutamicum CGMCC1.15647

ISSN: 1475-2859