Optimizing bulk segregant analysis of drug resistance using Plasmodium falciparum genetic crosses conducted in humanized mice
Issued Date
2022-04-15
Resource Type
eISSN
25890042
Scopus ID
2-s2.0-85127195874
Journal Title
iScience
Volume
25
Issue
4
Rights Holder(s)
SCOPUS
Bibliographic Citation
iScience Vol.25 No.4 (2022)
Suggested Citation
Brenneman K.V., Li X., Kumar S., Delgado E., Checkley L.A., Shoue D.A., Reyes A., Abatiyow B.A., Haile M.T., Tripura R., Peto T., Lek D., Button-Simons K.A., Kappe S.H.I., Dhorda M., Nosten F., Nkhoma S.C., Cheeseman I.H., Vaughan A.M., Ferdig M.T., Anderson T.J.C. Optimizing bulk segregant analysis of drug resistance using Plasmodium falciparum genetic crosses conducted in humanized mice. iScience Vol.25 No.4 (2022). doi:10.1016/j.isci.2022.104095 Retrieved from: https://repository.li.mahidol.ac.th/handle/20.500.14594/86525
Title
Optimizing bulk segregant analysis of drug resistance using Plasmodium falciparum genetic crosses conducted in humanized mice
Author's Affiliation
Faculty of Tropical Medicine, Mahidol University
National Institute of Public Health Cambodia
Texas Biomedical Research Institute
University of Notre Dame
University of Washington
American Type Culture Collection
Nuffield Department of Medicine
Seattle Biomedical Research Institute
National Center for Parasitology
National Institute of Public Health Cambodia
Texas Biomedical Research Institute
University of Notre Dame
University of Washington
American Type Culture Collection
Nuffield Department of Medicine
Seattle Biomedical Research Institute
National Center for Parasitology
Other Contributor(s)
Abstract
Classical malaria parasite genetic crosses involve isolation, genotyping, and phenotyping of progeny parasites, which is time consuming and laborious. We tested a rapid alternative approach—bulk segregant analysis (BSA)—that utilizes sequencing of bulk progeny populations with and without drug selection for rapid identification of drug resistance loci. We used dihydroartemisinin (DHA) selection in two genetic crosses and investigated how synchronization, cryopreservation, and the drug selection regimen impacted BSA success. We detected a robust quantitative trait locus (QTL) at kelch13 in both crosses but did not detect QTLs at four other candidate loci. QTLs were detected using synchronized, but not unsynchronized progeny pools, consistent with the stage-specific action of DHA. We also successfully applied BSA to cryopreserved progeny pools, expanding the utility of this approach. We conclude that BSA provides a powerful approach for investigating the genetic architecture of drug resistance in Plasmodium falciparum.
