Transforming the CRISPR/dCas9-based gene regulation technique into a forward screening tool in Plasmodium falciparum.

ABSTRACT: It is a significant challenge to assess the functions of many uncharacterized genes in human malaria parasites. Here, we present a genetic screening tool to assess the contribution of essential genes from Plasmodium falciparum by the conditional CRISPR-/deadCas9-based interference and activation (i/a) systems. We screened both CRISPRi and CRISPRa sets, consisting of nine parasite lines per set targeting nine genes via their respective gRNAs. By conducting amplicon sequencing of gRNA loci, we identified the contribution of each targeted gene to parasite fitness upon drug (artemisinin, chloroquine) and stress (starvation, heat shock) treatment. The screening was highly reproducible, and the screening libraries were easily generated by transfection of mixed plasmids expressing differen

SUBMITTER: Lucky AB 

PROVIDER: S-EPMC11015506 | biostudies-literature | 2024 Apr

REPOSITORIES: biostudies-literature