Study of C. glabrata infection in D. melanogaster after genome engineering by the CRISPR-Cas9 system.
(A) Scheme of Cas9-directed gene disruption in C. glabrata prior to D. melanogaster infection. After sequential transformation of sgRNA- and CAS9 (CYC1)-expressing vectors, targeted gene disruption was checked by sequencing. Vectors were then lost by growing yeasts overnight in liquid SCGlc 2% media. (B) Survival curves of immuno-competent A5001 flies and immuno-compromised MyD88 flies after infection with clean prick (cp), ∆HTL strain or ∆HTL + Cas9. Survival are shown as percentage of surviving flies. **p < 0,05 when compared to survival curve of MyD88 + cp or A5001 + cp. (C) Colony Forming Units (CFU) of cp and C. glabrata ∆HTL or ∆HTL + Cas9 after infection of D. melanogaster A5001 and MyD88 flies. (D) Infection of A5001 and MyD88 flies with the ∆HTL strain and strains disrupted for YPS11 and VPK1. ****p < 0,0001 when compared to survival cure of MyD88 + ∆HTL; ns, not significant. (E) CFU experiments related to infections performed in (D). Drosophila drawing is the work of B. Nuhanen, and was obtained from https://commons.wikimedia.org/wiki/File:Drosophila-drawing.svg, and is licensed under a CC BY-SA license (https://creativecommons.org/licenses/by-sa/3.0/deed.en). (F) Infection of latex (LTX) beads-injected A5001 and MyD88 flies with the ∆HTL strain or the NHEJ-vpk1 mutant.