CRISPR/Cas9 and genetic screens in malaria parasites: small genomes, big impact

T Ishizaki, S Hernandez, MS Paoletta… - Biochemical Society …, 2022 - portlandpress.com
T Ishizaki, S Hernandez, MS Paoletta, T Sanderson, ESC Bushell
Biochemical Society Transactions, 2022portlandpress.com
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 …
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.
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