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. 2021 Nov 11;220(12):e202104087. doi: 10.1083/jcb.202104087

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© 2021 Perez-Vale et al.

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Figure S4. — Generating a second cnoΔFAB allele directly in the cno locus. (A) Diagram of the cno genomic locus. Gray box outlines the region manipulated. gRNA1 targeted exon 18 and gRNA2 targeted a sequence downstream of the 3′ UTR. A dsDNA donor was provided for repair via homologous recombination. The dsDNA donor coding sequence is derived from cnoΔFAB-GFP with the gRNA1 PAM site mutated, and dsRed was used as a selectable marker. (B) Adult fly from stock homozygous for cnoΔFAB-GFP allele. (C) Immunoblot of embryonic extracts from 1–4 h, blotted with antibodies to Cno, GFP, and α-tubulin as a loading control. Magenta arrowhead indicates CnoΔRA.