Exploring the virulence gene interactome with CRISPR / dC as9 in the human malaria parasite
© 2020 The Authors. Published under the terms of the CC BY 4.0 license Mutually exclusive expression of the var multigene family is key to immune evasion and pathogenesis in Plasmodium falciparum, but few factors have been shown to play a direct role. We adapted a CRISPR-based proteomics approach to...
| Main Authors: | , , , , , , , , |
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| Format: | Article |
| Language: | English |
| Published: |
EMBO
2021
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| Online Access: | https://hdl.handle.net/1721.1/133513 |
| _version_ | 1811080408906858496 |
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| author | Bryant, Jessica M Baumgarten, Sebastian Dingli, Florent Loew, Damarys Sinha, Ameya Claës, Aurélie Preiser, Peter R Dedon, Peter C Scherf, Artur |
| author_facet | Bryant, Jessica M Baumgarten, Sebastian Dingli, Florent Loew, Damarys Sinha, Ameya Claës, Aurélie Preiser, Peter R Dedon, Peter C Scherf, Artur |
| author_sort | Bryant, Jessica M |
| collection | MIT |
| description | © 2020 The Authors. Published under the terms of the CC BY 4.0 license Mutually exclusive expression of the var multigene family is key to immune evasion and pathogenesis in Plasmodium falciparum, but few factors have been shown to play a direct role. We adapted a CRISPR-based proteomics approach to identify novel factors associated with var genes in their natural chromatin context. Catalytically inactive Cas9 (“dCas9”) was targeted to var gene regulatory elements, immunoprecipitated, and analyzed with mass spectrometry. Known and novel factors were enriched including structural proteins, DNA helicases, and chromatin remodelers. Functional characterization of PfISWI, an evolutionarily divergent putative chromatin remodeler enriched at the var gene promoter, revealed a role in transcriptional activation. Proteomics of PfISWI identified several proteins enriched at the var gene promoter such as acetyl-CoA synthetase, a putative MORC protein, and an ApiAP2 transcription factor. These findings validate the CRISPR/dCas9 proteomics method and define a new var gene-associated chromatin complex. This study establishes a tool for targeted chromatin purification of unaltered genomic loci and identifies novel chromatin-associated factors potentially involved in transcriptional control and/or chromatin organization of virulence genes in the human malaria parasite. |
| first_indexed | 2024-09-23T11:31:11Z |
| format | Article |
| id | mit-1721.1/133513 |
| institution | Massachusetts Institute of Technology |
| language | English |
| last_indexed | 2024-09-23T11:31:11Z |
| publishDate | 2021 |
| publisher | EMBO |
| record_format | dspace |
| spelling | mit-1721.1/1335132021-10-28T04:15:27Z Exploring the virulence gene interactome with CRISPR / dC as9 in the human malaria parasite Bryant, Jessica M Baumgarten, Sebastian Dingli, Florent Loew, Damarys Sinha, Ameya Claës, Aurélie Preiser, Peter R Dedon, Peter C Scherf, Artur © 2020 The Authors. Published under the terms of the CC BY 4.0 license Mutually exclusive expression of the var multigene family is key to immune evasion and pathogenesis in Plasmodium falciparum, but few factors have been shown to play a direct role. We adapted a CRISPR-based proteomics approach to identify novel factors associated with var genes in their natural chromatin context. Catalytically inactive Cas9 (“dCas9”) was targeted to var gene regulatory elements, immunoprecipitated, and analyzed with mass spectrometry. Known and novel factors were enriched including structural proteins, DNA helicases, and chromatin remodelers. Functional characterization of PfISWI, an evolutionarily divergent putative chromatin remodeler enriched at the var gene promoter, revealed a role in transcriptional activation. Proteomics of PfISWI identified several proteins enriched at the var gene promoter such as acetyl-CoA synthetase, a putative MORC protein, and an ApiAP2 transcription factor. These findings validate the CRISPR/dCas9 proteomics method and define a new var gene-associated chromatin complex. This study establishes a tool for targeted chromatin purification of unaltered genomic loci and identifies novel chromatin-associated factors potentially involved in transcriptional control and/or chromatin organization of virulence genes in the human malaria parasite. 2021-10-27T19:53:16Z 2021-10-27T19:53:16Z 2020 2021-08-26T15:54:10Z Article http://purl.org/eprint/type/JournalArticle https://hdl.handle.net/1721.1/133513 en 10.15252/MSB.20209569 Molecular Systems Biology Creative Commons Attribution 4.0 International license https://creativecommons.org/licenses/by/4.0/ application/pdf EMBO EMBO Press |
| spellingShingle | Bryant, Jessica M Baumgarten, Sebastian Dingli, Florent Loew, Damarys Sinha, Ameya Claës, Aurélie Preiser, Peter R Dedon, Peter C Scherf, Artur Exploring the virulence gene interactome with CRISPR / dC as9 in the human malaria parasite |
| title | Exploring the virulence gene interactome with CRISPR / dC as9 in the human malaria parasite |
| title_full | Exploring the virulence gene interactome with CRISPR / dC as9 in the human malaria parasite |
| title_fullStr | Exploring the virulence gene interactome with CRISPR / dC as9 in the human malaria parasite |
| title_full_unstemmed | Exploring the virulence gene interactome with CRISPR / dC as9 in the human malaria parasite |
| title_short | Exploring the virulence gene interactome with CRISPR / dC as9 in the human malaria parasite |
| title_sort | exploring the virulence gene interactome with crispr dc as9 in the human malaria parasite |
| url | https://hdl.handle.net/1721.1/133513 |
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