CRISPR/Cas9 and genetic screens in malaria parasites : small genomes, big impact
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. I...
| Autores principales: | , , , , |
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| Formato: | Artículo |
| Lenguaje: | Inglés |
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Portland Press
2022
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| Acceso en línea: | http://hdl.handle.net/20.500.12123/12437 https://portlandpress.com/biochemsoctrans/article/50/3/1069/231360/CRISPR-Cas9-and-genetic-screens-in-malaria https://doi.org/10.1042/BST20210281 |
| _version_ | 1855484973202538496 |
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| author | Ishizaki, Takahiro Hernandez, Sophia Paoletta, Martina Sanderson, Theo Bushell, Ellen S. C. |
| author_browse | Bushell, Ellen S. C. Hernandez, Sophia Ishizaki, Takahiro Paoletta, Martina Sanderson, Theo |
| author_facet | Ishizaki, Takahiro Hernandez, Sophia Paoletta, Martina Sanderson, Theo Bushell, Ellen S. C. |
| author_sort | Ishizaki, Takahiro |
| collection | INTA Digital |
| description | 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. |
| format | Artículo |
| id | INTA12437 |
| institution | Instituto Nacional de Tecnología Agropecuaria (INTA -Argentina) |
| language | Inglés |
| publishDate | 2022 |
| publishDateRange | 2022 |
| publishDateSort | 2022 |
| publisher | Portland Press |
| publisherStr | Portland Press |
| record_format | dspace |
| spelling | INTA124372022-07-29T11:11:05Z CRISPR/Cas9 and genetic screens in malaria parasites : small genomes, big impact Ishizaki, Takahiro Hernandez, Sophia Paoletta, Martina Sanderson, Theo Bushell, Ellen S. C. Molecular Genetics Laboratory Techniques CRISPR Parasitology Genomes Genética Molecular Técnicas de Laboratorio Repeticiones Palindrómicas Cortas Agrupadas y Regularmente Interespaciadas Malaria Plasmodium falciparum Parasitología Genomas Host–microbe Interactions Interacciones Huésped-microbio 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. Instituto de Biotecnología Fil: Ishizaki, Takahiro. Umeå University. Department of Molecular Biology; Suecia Fil: Ishizaki, Takahiro. The Laboratory for Molecular Infection Medicine Sweden (MIMS); Suecia Fil: Hernandez, Sophia. Umeå University. Department of Molecular Biology; Suecia Fil: Hernandez, Sophia. The Laboratory for Molecular Infection Medicine Sweden (MIMS); Suecia Fil: Paoletta, Martina. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Agrobiotecnología y Biología Molecular; Argentina Fil: Paoletta, Martina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina Fil: Paoletta, Martina. Umeå University. Department of Molecular Biology; Suecia Fil: Paoletta, Martina. The Laboratory for Molecular Infection Medicine Sweden (MIMS); Suecia Fil: Sanderson, Theo. Francis Crick Institute; Reino Unido Fil: Bushell, Ellen S. C. Umeå University. Department of Molecular Biology; Suecia Fil: Bushell, Ellen S. C. The Laboratory for Molecular Infection Medicine Sweden (MIMS); Suecia 2022-07-29T10:52:58Z 2022-07-29T10:52:58Z 2022-06 info:ar-repo/semantics/artículo info:eu-repo/semantics/article info:eu-repo/semantics/publishedVersion http://hdl.handle.net/20.500.12123/12437 https://portlandpress.com/biochemsoctrans/article/50/3/1069/231360/CRISPR-Cas9-and-genetic-screens-in-malaria 1470-8752 https://doi.org/10.1042/BST20210281 eng info:eu-repograntAgreement/INTA/2019-PD-E5-I105-001/2019-PD-E5-I105-001/AR./Patógenos animales: su interacción con el hospedador y el medio ambiente. Impacto en productividad, ecosistemas, sanidad animal y salud pública en el marco “Una Salud” info:eu-repo/semantics/openAccess http://creativecommons.org/licenses/by-nc-sa/4.0/ Creative Commons Attribution-NonCommercial-ShareAlike 4.0 International (CC BY-NC-SA 4.0) application/pdf Portland Press Biochemical Society Transactions 50 (3) : 1069-1079 (Junio 2022) |
| spellingShingle | Molecular Genetics Laboratory Techniques CRISPR Parasitology Genomes Genética Molecular Técnicas de Laboratorio Repeticiones Palindrómicas Cortas Agrupadas y Regularmente Interespaciadas Malaria Plasmodium falciparum Parasitología Genomas Host–microbe Interactions Interacciones Huésped-microbio Ishizaki, Takahiro Hernandez, Sophia Paoletta, Martina Sanderson, Theo Bushell, Ellen S. C. CRISPR/Cas9 and genetic screens in malaria parasites : small genomes, big impact |
| title | CRISPR/Cas9 and genetic screens in malaria parasites : small genomes, big impact |
| title_full | CRISPR/Cas9 and genetic screens in malaria parasites : small genomes, big impact |
| title_fullStr | CRISPR/Cas9 and genetic screens in malaria parasites : small genomes, big impact |
| title_full_unstemmed | CRISPR/Cas9 and genetic screens in malaria parasites : small genomes, big impact |
| title_short | CRISPR/Cas9 and genetic screens in malaria parasites : small genomes, big impact |
| title_sort | crispr cas9 and genetic screens in malaria parasites small genomes big impact |
| topic | Molecular Genetics Laboratory Techniques CRISPR Parasitology Genomes Genética Molecular Técnicas de Laboratorio Repeticiones Palindrómicas Cortas Agrupadas y Regularmente Interespaciadas Malaria Plasmodium falciparum Parasitología Genomas Host–microbe Interactions Interacciones Huésped-microbio |
| url | http://hdl.handle.net/20.500.12123/12437 https://portlandpress.com/biochemsoctrans/article/50/3/1069/231360/CRISPR-Cas9-and-genetic-screens-in-malaria https://doi.org/10.1042/BST20210281 |
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