An integrated platform for genome engineering and gene expression perturbation in Plasmodium falciparum
Abstract Establishing robust genome engineering methods in the malarial parasite, Plasmodium falciparum, has the potential to substantially improve the efficiency with which we gain understanding of this pathogen’s biology to propel treatment and elimination efforts. Methods for manipulating gene ex...
| Main Authors: | , , , , , , , |
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| Format: | Article |
| Language: | English |
| Published: |
Nature Portfolio
2021-01-01
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| Series: | Scientific Reports |
| Online Access: | https://doi.org/10.1038/s41598-020-77644-4 |
| _version_ | 1818840196918018048 |
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| author | Armiyaw S. Nasamu Alejandra Falla Charisse Flerida A. Pasaje Bridget A. Wall Jeffrey C. Wagner Suresh M. Ganesan Stephen J. Goldfless Jacquin C. Niles |
| author_facet | Armiyaw S. Nasamu Alejandra Falla Charisse Flerida A. Pasaje Bridget A. Wall Jeffrey C. Wagner Suresh M. Ganesan Stephen J. Goldfless Jacquin C. Niles |
| author_sort | Armiyaw S. Nasamu |
| collection | DOAJ |
| description | Abstract Establishing robust genome engineering methods in the malarial parasite, Plasmodium falciparum, has the potential to substantially improve the efficiency with which we gain understanding of this pathogen’s biology to propel treatment and elimination efforts. Methods for manipulating gene expression and engineering the P. falciparum genome have been validated. However, a significant barrier to fully leveraging these advances is the difficulty associated with assembling the extremely high AT content DNA constructs required for modifying the P. falciparum genome. These are frequently unstable in commonly-used circular plasmids. We address this bottleneck by devising a DNA assembly framework leveraging the improved reliability with which large AT-rich regions can be efficiently manipulated in linear plasmids. This framework integrates several key functional genetics outcomes via CRISPR/Cas9 and other methods from a common, validated framework. Overall, this molecular toolkit enables P. falciparum genetics broadly and facilitates deeper interrogation of parasite genes involved in diverse biological processes. |
| first_indexed | 2024-12-19T04:06:21Z |
| format | Article |
| id | doaj.art-e9d76cd561ad4d1a993ae54780056e38 |
| institution | Directory Open Access Journal |
| issn | 2045-2322 |
| language | English |
| last_indexed | 2024-12-19T04:06:21Z |
| publishDate | 2021-01-01 |
| publisher | Nature Portfolio |
| record_format | Article |
| series | Scientific Reports |
| spelling | doaj.art-e9d76cd561ad4d1a993ae54780056e382022-12-21T20:36:32ZengNature PortfolioScientific Reports2045-23222021-01-0111111510.1038/s41598-020-77644-4An integrated platform for genome engineering and gene expression perturbation in Plasmodium falciparumArmiyaw S. Nasamu0Alejandra Falla1Charisse Flerida A. Pasaje2Bridget A. Wall3Jeffrey C. Wagner4Suresh M. Ganesan5Stephen J. Goldfless6Jacquin C. Niles7Department of Biological Engineering, Massachusetts Institute of TechnologyDepartment of Biological Engineering, Massachusetts Institute of TechnologyDepartment of Biological Engineering, Massachusetts Institute of TechnologyDepartment of Biological Engineering, Massachusetts Institute of TechnologyDepartment of Biological Engineering, Massachusetts Institute of TechnologyDepartment of Biological Engineering, Massachusetts Institute of TechnologyDepartment of Biological Engineering, Massachusetts Institute of TechnologyDepartment of Biological Engineering, Massachusetts Institute of TechnologyAbstract Establishing robust genome engineering methods in the malarial parasite, Plasmodium falciparum, has the potential to substantially improve the efficiency with which we gain understanding of this pathogen’s biology to propel treatment and elimination efforts. Methods for manipulating gene expression and engineering the P. falciparum genome have been validated. However, a significant barrier to fully leveraging these advances is the difficulty associated with assembling the extremely high AT content DNA constructs required for modifying the P. falciparum genome. These are frequently unstable in commonly-used circular plasmids. We address this bottleneck by devising a DNA assembly framework leveraging the improved reliability with which large AT-rich regions can be efficiently manipulated in linear plasmids. This framework integrates several key functional genetics outcomes via CRISPR/Cas9 and other methods from a common, validated framework. Overall, this molecular toolkit enables P. falciparum genetics broadly and facilitates deeper interrogation of parasite genes involved in diverse biological processes.https://doi.org/10.1038/s41598-020-77644-4 |
| spellingShingle | Armiyaw S. Nasamu Alejandra Falla Charisse Flerida A. Pasaje Bridget A. Wall Jeffrey C. Wagner Suresh M. Ganesan Stephen J. Goldfless Jacquin C. Niles An integrated platform for genome engineering and gene expression perturbation in Plasmodium falciparum Scientific Reports |
| title | An integrated platform for genome engineering and gene expression perturbation in Plasmodium falciparum |
| title_full | An integrated platform for genome engineering and gene expression perturbation in Plasmodium falciparum |
| title_fullStr | An integrated platform for genome engineering and gene expression perturbation in Plasmodium falciparum |
| title_full_unstemmed | An integrated platform for genome engineering and gene expression perturbation in Plasmodium falciparum |
| title_short | An integrated platform for genome engineering and gene expression perturbation in Plasmodium falciparum |
| title_sort | integrated platform for genome engineering and gene expression perturbation in plasmodium falciparum |
| url | https://doi.org/10.1038/s41598-020-77644-4 |
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