CRISPR-Cas9 high-throughput screening to study drug resistance in Leishmania infantum.

ABSTRACT: Parasites of the genus Leishmania pose a global health threat with limited treatment options. New drugs are urgently needed, and genomic screens have the potential to accelerate target discovery, mode of action, and resistance mechanisms against these new drugs. We describe here our effort in developing a genome-wide CRISPR-Cas9 screen in Leishmania, an organism lacking a functional nonhomologous end joining system that must rely on microhomology-mediated end joining, single-strand annealing, or homologous recombination for repairing Cas9-induced double-stranded DNA breaks. A new vector for cloning and expressing single guide RNAs (sgRNAs) was designed and proven to be effective in a small pilot project while enriching specific sgRNAs during drug selection. We then developed

SUBMITTER: Queffeulou M 

PROVIDER: S-EPMC11253630 | biostudies-literature | 2024 Jul

REPOSITORIES: biostudies-literature