CRISPR/Cas9 and genetic screens in malaria parasites: small genomes, big impactShow others and affiliations
2022 (English)In: Biochemical Society Transactions, ISSN 0300-5127, E-ISSN 1470-8752, Vol. 50, no 3, p. 1069-1079Article, review/survey (Refereed) Published
Abstract [en]
The ∼30 Mb genomes of the Plasmodium parasites that cause malaria each encode ∼5000 genes, but the functions of the majority remain unknown. This is due to a paucity of functional annotation from sequence homology, which is compounded by low genetic tractability compared with many model organisms. In recent years technical breakthroughs have made forward and reverse genome-scale screens in Plasmodium possible. Furthermore, the adaptation of Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR) and CRISPR-Associated protein 9 (CRISPR/Cas9) technology has dramatically improved gene editing efficiency at the single gene level. Here, we review the arrival of genetic screens in malaria parasites to analyse parasite gene function at a genome-scale and their impact on understanding parasite biology. CRISPR/Cas9 screens, which have revolutionised human and model organism research, have not yet been implemented in malaria parasites due to the need for more complex CRISPR/Cas9 gene targeting vector libraries. We therefore introduce the reader to CRISPR-based screens in the related apicomplexan Toxoplasma gondii and discuss how these approaches could be adapted to develop CRISPR/Cas9 based genome-scale genetic screens in malaria parasites. Moreover, since more than half of Plasmodium genes are required for normal asexual blood-stage reproduction, and cannot be targeted using knockout methods, we discuss how CRISPR/Cas9 could be used to scale up conditional gene knockdown approaches to systematically assign function to essential genes.
Place, publisher, year, edition, pages
Portland Press, 2022. Vol. 50, no 3, p. 1069-1079
Keywords [en]
Plasmodium falciparum, biochemical techniques and resources, CRISPR, genetics, malaria
National Category
Cell and Molecular Biology Medical Biotechnology (with a focus on Cell Biology (including Stem Cell Biology), Molecular Biology, Microbiology, Biochemistry or Biopharmacy)
Identifiers
URN: urn:nbn:se:umu:diva-197999DOI: 10.1042/BST20210281ISI: 000804375700001PubMedID: 35621119Scopus ID: 2-s2.0-85133214886OAI: oai:DiVA.org:umu-197999DiVA, id: diva2:1682676
Funder
Swedish Research Council, 2021-06602Knut and Alice Wallenberg Foundation, 2019.0178Wellcome trust, 210918/Z/18/Z2022-07-112022-07-112022-07-11Bibliographically approved