Efficient genome editing with CRISPR/Cas9 in Pleurotus ostreatus

AMB Express

Table 3 Comparison of transformants and mutation frequency obtained using CRISPR/Cas9 and donor DNA templates

Plasmid with sgRNA	Donor DNA template	Hygromycin-resistant strains				5-FC-resistant strains				fcy1 mutant by PCR^a
Plasmid with sgRNA	Donor DNA template	Rep 1	Rep 2	Rep 3	Total	Rep 1	Rep 2	Rep 3	Total	Rep 1	Rep 2	Rep 3	Total
fcy1 sgRNA No. 2	–	0	2	2	4	0	0	0	0	0	0	0	0
	0.2 kb	0	2	2	4	0	1	1	2 (50%)^b	0	0	0	0
	0.5 kb	4	5	4	13	4	3	3	10 (76.9%)^b	4	2	3	9 (69.2%)^c
	1 kb	2	2	2	6	2	1	2	5 (83.3%)^b	2	1	2	5 (83.3%)^c

^aThe number of mutants which can be identified by difference size, or no amplified, bands compared to wild-type control were observed in the genomic PCR experiment (Additional file 1: Fig. S2C)
^bPercentage of 5-FC-resistant strains from the total number of hygromycin-resistant strains
^cPercentage of fcy1 mutants from the total number of hygromycin-resistant strains