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Large DNA fragment knock-in and sequential gene editing in Plasmodium falciparum: a preliminary study using suicide-rescue-based CRISPR/Cas9 system
CRISPR/Cas9 technology applied to Plasmodium falciparum offers the potential to greatly improve gene editing, but such expectations including large DNA fragment knock-ins and sequential gene editing have remained unfulfilled. Here, we achieved a major advance in addressing this challenge, especially...
Autores principales: | , , , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Springer US
2023
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10066980/ https://www.ncbi.nlm.nih.gov/pubmed/37004637 http://dx.doi.org/10.1007/s11010-023-04711-5 |
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author | Lu, Junnan Tong, Ying Dong, Rui Yang, Yijun Hu, Wen Zhang, Minghong Liu, Quan Zhao, Siting Adams, John H. Qin, Li Chen, Xiaoping |
author_facet | Lu, Junnan Tong, Ying Dong, Rui Yang, Yijun Hu, Wen Zhang, Minghong Liu, Quan Zhao, Siting Adams, John H. Qin, Li Chen, Xiaoping |
author_sort | Lu, Junnan |
collection | PubMed |
description | CRISPR/Cas9 technology applied to Plasmodium falciparum offers the potential to greatly improve gene editing, but such expectations including large DNA fragment knock-ins and sequential gene editing have remained unfulfilled. Here, we achieved a major advance in addressing this challenge, especially for creating large DNA fragment knock-ins and sequential editing, by modifying our suicide-rescue-based system that has already been demonstrated to be highly efficient for conventional gene editing. This improved approach was confirmed to mediate efficient knock-ins of DNA fragments up to 6.3 kb, to produce “marker-free” genetically engineered parasites and to show potential for sequential gene editing. This represents an important advancement in establishing platforms for large-scale genome editing, which might gain a better understanding of gene function for the most lethal cause of malaria and contribute to adjusting synthetic biology strategies to live parasite malaria vaccine development. Site-directed knock-in of large DNA fragments is highly efficient using suicide-rescue-based CRISPR/Cas9 system, and sequential gene insertion is feasible but further confirmation is still needed. SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1007/s11010-023-04711-5. |
format | Online Article Text |
id | pubmed-10066980 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2023 |
publisher | Springer US |
record_format | MEDLINE/PubMed |
spelling | pubmed-100669802023-04-03 Large DNA fragment knock-in and sequential gene editing in Plasmodium falciparum: a preliminary study using suicide-rescue-based CRISPR/Cas9 system Lu, Junnan Tong, Ying Dong, Rui Yang, Yijun Hu, Wen Zhang, Minghong Liu, Quan Zhao, Siting Adams, John H. Qin, Li Chen, Xiaoping Mol Cell Biochem Article CRISPR/Cas9 technology applied to Plasmodium falciparum offers the potential to greatly improve gene editing, but such expectations including large DNA fragment knock-ins and sequential gene editing have remained unfulfilled. Here, we achieved a major advance in addressing this challenge, especially for creating large DNA fragment knock-ins and sequential editing, by modifying our suicide-rescue-based system that has already been demonstrated to be highly efficient for conventional gene editing. This improved approach was confirmed to mediate efficient knock-ins of DNA fragments up to 6.3 kb, to produce “marker-free” genetically engineered parasites and to show potential for sequential gene editing. This represents an important advancement in establishing platforms for large-scale genome editing, which might gain a better understanding of gene function for the most lethal cause of malaria and contribute to adjusting synthetic biology strategies to live parasite malaria vaccine development. Site-directed knock-in of large DNA fragments is highly efficient using suicide-rescue-based CRISPR/Cas9 system, and sequential gene insertion is feasible but further confirmation is still needed. SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1007/s11010-023-04711-5. Springer US 2023-04-01 /pmc/articles/PMC10066980/ /pubmed/37004637 http://dx.doi.org/10.1007/s11010-023-04711-5 Text en © The Author(s), under exclusive licence to Springer Science+Business Media, LLC, part of Springer Nature 2023, Springer Nature or its licensor (e.g. a society or other partner) holds exclusive rights to this article under a publishing agreement with the author(s) or other rightsholder(s); author self-archiving of the accepted manuscript version of this article is solely governed by the terms of such publishing agreement and applicable law. This article is made available via the PMC Open Access Subset for unrestricted research re-use and secondary analysis in any form or by any means with acknowledgement of the original source. These permissions are granted for the duration of the World Health Organization (WHO) declaration of COVID-19 as a global pandemic. |
spellingShingle | Article Lu, Junnan Tong, Ying Dong, Rui Yang, Yijun Hu, Wen Zhang, Minghong Liu, Quan Zhao, Siting Adams, John H. Qin, Li Chen, Xiaoping Large DNA fragment knock-in and sequential gene editing in Plasmodium falciparum: a preliminary study using suicide-rescue-based CRISPR/Cas9 system |
title | Large DNA fragment knock-in and sequential gene editing in Plasmodium falciparum: a preliminary study using suicide-rescue-based CRISPR/Cas9 system |
title_full | Large DNA fragment knock-in and sequential gene editing in Plasmodium falciparum: a preliminary study using suicide-rescue-based CRISPR/Cas9 system |
title_fullStr | Large DNA fragment knock-in and sequential gene editing in Plasmodium falciparum: a preliminary study using suicide-rescue-based CRISPR/Cas9 system |
title_full_unstemmed | Large DNA fragment knock-in and sequential gene editing in Plasmodium falciparum: a preliminary study using suicide-rescue-based CRISPR/Cas9 system |
title_short | Large DNA fragment knock-in and sequential gene editing in Plasmodium falciparum: a preliminary study using suicide-rescue-based CRISPR/Cas9 system |
title_sort | large dna fragment knock-in and sequential gene editing in plasmodium falciparum: a preliminary study using suicide-rescue-based crispr/cas9 system |
topic | Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10066980/ https://www.ncbi.nlm.nih.gov/pubmed/37004637 http://dx.doi.org/10.1007/s11010-023-04711-5 |
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