Precise single base substitution in the shibire gene by CRISPR/Cas9-mediated homology directed repair in Bactrocera tryoni

Table 1 Summary of the microinjections performed using the crRNA-tracrRNA RNP complex system to introduce the shi^ts1 point mutation into B. tryoni

Injections	Concentration of ssODN	# embryos injected	# G₀ adults (% survival)	# successful G₀ matings	# G₀ germline mutants^a	Detected mutation
Injections	Concentration of ssODN	# embryos injected	# G₀ adults (% survival)	# successful G₀ matings	# G₀ germline mutants^a	Mutation	# G₁ mutants^b
Shi^ts1	200 ng/μL	254	14 (5.5%)	7	2 (14.3%)	Del (4 bp)	7 out of 12
	200 ng/μL	254	14 (5.5%)	7	2 (14.3%)	KI (Shi^ts1)	18 out of 120
	250 ng/μL	473	10 (2.1%)	8	0	0	0
	300 ng/μL	310	5 (1.6%)	2	0	0	0

Del Deletion, KI Knock-in
^aThe mutagenesis efficiency is presented in brackets as the percentage of G₀ flies with an identified germline mutation out of the total number of G₀ adults obtained
^bThe number of G₁ mutants identified out of a total of G₁ progeny screened for that particular G₀ germline mutant

ISSN: 2730-6844