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Rapid and iterative genome editing in the malaria parasite Plasmodium knowlesi provides new tools for P. vivax research
Tackling relapsing Plasmodium vivax and zoonotic Plasmodium knowlesi infections is critical to reducing malaria incidence and mortality worldwide. Understanding the biology of these important and related parasites was previously constrained by the lack of robust molecular and genetic approaches. Her...
Autores principales: | , , , , , , , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
eLife Sciences Publications, Ltd
2019
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6579517/ https://www.ncbi.nlm.nih.gov/pubmed/31205002 http://dx.doi.org/10.7554/eLife.45829 |
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author | Mohring, Franziska Hart, Melissa Natalie Rawlinson, Thomas A Henrici, Ryan Charleston, James A Diez Benavente, Ernest Patel, Avnish Hall, Joanna Almond, Neil Campino, Susana Clark, Taane G Sutherland, Colin J Baker, David A Draper, Simon J Moon, Robert William |
author_facet | Mohring, Franziska Hart, Melissa Natalie Rawlinson, Thomas A Henrici, Ryan Charleston, James A Diez Benavente, Ernest Patel, Avnish Hall, Joanna Almond, Neil Campino, Susana Clark, Taane G Sutherland, Colin J Baker, David A Draper, Simon J Moon, Robert William |
author_sort | Mohring, Franziska |
collection | PubMed |
description | Tackling relapsing Plasmodium vivax and zoonotic Plasmodium knowlesi infections is critical to reducing malaria incidence and mortality worldwide. Understanding the biology of these important and related parasites was previously constrained by the lack of robust molecular and genetic approaches. Here, we establish CRISPR-Cas9 genome editing in a culture-adapted P. knowlesi strain and define parameters for optimal homology-driven repair. We establish a scalable protocol for the production of repair templates by PCR and demonstrate the flexibility of the system by tagging proteins with distinct cellular localisations. Using iterative rounds of genome-editing we generate a transgenic line expressing P. vivax Duffy binding protein (PvDBP), a lead vaccine candidate. We demonstrate that PvDBP plays no role in reticulocyte restriction but can alter the macaque/human host cell tropism of P. knowlesi. Critically, antibodies raised against the P. vivax antigen potently inhibit proliferation of this strain, providing an invaluable tool to support vaccine development. |
format | Online Article Text |
id | pubmed-6579517 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2019 |
publisher | eLife Sciences Publications, Ltd |
record_format | MEDLINE/PubMed |
spelling | pubmed-65795172019-06-19 Rapid and iterative genome editing in the malaria parasite Plasmodium knowlesi provides new tools for P. vivax research Mohring, Franziska Hart, Melissa Natalie Rawlinson, Thomas A Henrici, Ryan Charleston, James A Diez Benavente, Ernest Patel, Avnish Hall, Joanna Almond, Neil Campino, Susana Clark, Taane G Sutherland, Colin J Baker, David A Draper, Simon J Moon, Robert William eLife Genetics and Genomics Tackling relapsing Plasmodium vivax and zoonotic Plasmodium knowlesi infections is critical to reducing malaria incidence and mortality worldwide. Understanding the biology of these important and related parasites was previously constrained by the lack of robust molecular and genetic approaches. Here, we establish CRISPR-Cas9 genome editing in a culture-adapted P. knowlesi strain and define parameters for optimal homology-driven repair. We establish a scalable protocol for the production of repair templates by PCR and demonstrate the flexibility of the system by tagging proteins with distinct cellular localisations. Using iterative rounds of genome-editing we generate a transgenic line expressing P. vivax Duffy binding protein (PvDBP), a lead vaccine candidate. We demonstrate that PvDBP plays no role in reticulocyte restriction but can alter the macaque/human host cell tropism of P. knowlesi. Critically, antibodies raised against the P. vivax antigen potently inhibit proliferation of this strain, providing an invaluable tool to support vaccine development. eLife Sciences Publications, Ltd 2019-06-17 /pmc/articles/PMC6579517/ /pubmed/31205002 http://dx.doi.org/10.7554/eLife.45829 Text en © 2019, Mohring et al http://creativecommons.org/licenses/by/4.0/ http://creativecommons.org/licenses/by/4.0/This article is distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0/) , which permits unrestricted use and redistribution provided that the original author and source are credited. |
spellingShingle | Genetics and Genomics Mohring, Franziska Hart, Melissa Natalie Rawlinson, Thomas A Henrici, Ryan Charleston, James A Diez Benavente, Ernest Patel, Avnish Hall, Joanna Almond, Neil Campino, Susana Clark, Taane G Sutherland, Colin J Baker, David A Draper, Simon J Moon, Robert William Rapid and iterative genome editing in the malaria parasite Plasmodium knowlesi provides new tools for P. vivax research |
title | Rapid and iterative genome editing in the malaria parasite Plasmodium knowlesi provides new tools for P. vivax research |
title_full | Rapid and iterative genome editing in the malaria parasite Plasmodium knowlesi provides new tools for P. vivax research |
title_fullStr | Rapid and iterative genome editing in the malaria parasite Plasmodium knowlesi provides new tools for P. vivax research |
title_full_unstemmed | Rapid and iterative genome editing in the malaria parasite Plasmodium knowlesi provides new tools for P. vivax research |
title_short | Rapid and iterative genome editing in the malaria parasite Plasmodium knowlesi provides new tools for P. vivax research |
title_sort | rapid and iterative genome editing in the malaria parasite plasmodium knowlesi provides new tools for p. vivax research |
topic | Genetics and Genomics |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6579517/ https://www.ncbi.nlm.nih.gov/pubmed/31205002 http://dx.doi.org/10.7554/eLife.45829 |
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