Correction of the auditory phenotype in C57BL/6N mice via CRISPR/Cas9-mediated homology directed repair

Table 1 Number of injected embryos, embryos transferred, pups born, and mutation rate

Design	Cas9 (D10A) mRNA [conc] ng/μl	sgRNAs [conc] ng/μl (each)	ssODN [conc] ng/μl	Number of injected embryos	Lysed after injecting	Lysed (%)	Number of ET	Number of pups born	Born/injected (%)	Number of TG mice	Mutation rate (%)	Number of correct F₀ mice	Legitimate repair rate (%)
1	100	50	20	24	3	12.5	21	11	45.8	1	9.1	0	0
1	200	100	40	220	20	9.1	200	61	27.7	10	16.4	2	3.3
2	100	50	20	101	11	10.9	90	16	15.8	0	0.0	0	0
2	200	100	40	111	4	3.6	107	16	14.4	4	25.0	2	12.5
				456	38	8.3	418	104	22.8	15	14.4	4	3.8

ET embryos transferred, TG transgenic
Generation of mice carrying the Cdh23 ^753A>G repair using pronuclear injection of CRISPR/Cas9 reagents. For designs 1 and 2, two sets of injections were performed using either 100, 50, 50 and 20 ng/μl or 200, 100, 100 and 40 ng/μl of Cas9 (D10A) nickase mRNA, sgRNAs_U, sgRNA_D and ssODN, respectively. Higher efficiency was obtained when using the higher concentrations. The percentages of mutation rate and legitimate repair rate have both been calculated using the number of pups born as the denominator

ISSN: 1756-994X