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CRISPR Editing

In addition to our genetic crosses, we also perform CRISPR/Cas9 editing on genes of interest that have been identified through BSA or phenotyping of progeny clones. Through our isogenic CRISPR lines, we have measured the fitness costs of various kelch13 mutations as well as the impact of different pfaat1 mutations on parasite fitness and resistance to frontline antimalarials.

Publications

Amambua-Ngwa A, Button-Simons KA, Li X, Kumar S, Brenneman KV, Ferrari M, Checkley LA, Haile MT, Shoue DA, McDew-White M, Tindall SM, Reyes A, Delgado E, Dalhoff H, Larbalestier JK, Amato R, Pearson RD, Taylor AB, Nosten FH, D'Alessandro U, Kwiatkowski D, Cheeseman IH, Kappe SHI, Avery SV, Conway DJ, Vaughan AM, Ferdig MT, Anderson TJC. Chloroquine resistance evolution in Plasmodium falciparum is mediated by the putative amino acid transporter AAT1. Nat Microbiol. 2023 Jul;8(7):1213-1226. doi: 10.1038/s41564-023-01377-z. Epub 2023 May 11. PMID: 37169919; PMCID: PMC10322710.

 

 

Diagram illustrating the relationships between different NHP4302 variants. Central circle labeled
Three panels labeled a, b, and c show data related to the pfaat1 genotype. Panel A diagrams four pfaat1 genotypes, NHP 4026, 258L/313S, 258L/313F, 258S/313S, and 258S/313F. Panel B shows a dot plot of CQ IC50 (nM) for the last four genotypes with p-values displayed above the data. Panel C shows a bar graph comparing the relative fitness of those same genotypes with p-values displayed above the graph.