Fig. 2

Experimental design to produce G₁ XX males bearing heteroallelic mutation of Cctra. Two sets of parallel Cas9-RNP injections to target Cctra into XX embryos can lead to both G₀ XX males and XX females. The G₀ adults from the two sets can bear some Cctra monoallelic independent mutations (mosaicism) both in the soma and in the germ line. In set n. 1, the injection mix contains also Cctra-dsRNA to induce full masculinization of XX embryos (Pane et al., 2002). Hence, the G₀ XX males from set n. 1 can be crossed with G₀ XX females from set n. 2, to produce a G₁ XX progeny, expected to be composed mostly of XX females (carrying either none or monoallelic mutations) and of few G₁ XX males, carrying heteroallelic mutations of Cctra (Cctra¹⁻/Cctra²⁻) inherited from the respective parents