A novel 'gene insertion/marker out' (GIMO) method for transgene expression and gene complementation in rodent malaria parasites.
Research on the biology of malaria parasites has greatly benefited from the application of reverse genetic technologies, in particular through the analysis of gene deletion mutants and studies on transgenic parasites that express heterologous or mutated proteins. However, transfection in Plasmodium...
| Main Authors: | , , , , , , , , |
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| Format: | Article |
| Language: | English |
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Public Library of Science (PLoS)
2011-01-01
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| Series: | PLoS ONE |
| Online Access: | https://www.ncbi.nlm.nih.gov/pmc/articles/pmid/22216235/pdf/?tool=EBI |
| _version_ | 1818341744399351808 |
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| author | Jing-wen Lin Takeshi Annoura Mohammed Sajid Séverine Chevalley-Maurel Jai Ramesar Onny Klop Blandine M D Franke-Fayard Chris J Janse Shahid M Khan |
| author_facet | Jing-wen Lin Takeshi Annoura Mohammed Sajid Séverine Chevalley-Maurel Jai Ramesar Onny Klop Blandine M D Franke-Fayard Chris J Janse Shahid M Khan |
| author_sort | Jing-wen Lin |
| collection | DOAJ |
| description | Research on the biology of malaria parasites has greatly benefited from the application of reverse genetic technologies, in particular through the analysis of gene deletion mutants and studies on transgenic parasites that express heterologous or mutated proteins. However, transfection in Plasmodium is limited by the paucity of drug-selectable markers that hampers subsequent genetic modification of the same mutant. We report the development of a novel 'gene insertion/marker out' (GIMO) method for two rodent malaria parasites, which uses negative selection to rapidly generate transgenic mutants ready for subsequent modifications. We have created reference mother lines for both P. berghei ANKA and P. yoelii 17XNL that serve as recipient parasites for GIMO-transfection. Compared to existing protocols GIMO-transfection greatly simplifies and speeds up the generation of mutants expressing heterologous proteins, free of drug-resistance genes, and requires far fewer laboratory animals. In addition we demonstrate that GIMO-transfection is also a simple and fast method for genetic complementation of mutants with a gene deletion or mutation. The implementation of GIMO-transfection procedures should greatly enhance Plasmodium reverse-genetic research. |
| first_indexed | 2024-12-13T16:03:40Z |
| format | Article |
| id | doaj.art-9f23ec21c36444b8b6062e927838ab68 |
| institution | Directory Open Access Journal |
| issn | 1932-6203 |
| language | English |
| last_indexed | 2024-12-13T16:03:40Z |
| publishDate | 2011-01-01 |
| publisher | Public Library of Science (PLoS) |
| record_format | Article |
| series | PLoS ONE |
| spelling | doaj.art-9f23ec21c36444b8b6062e927838ab682022-12-21T23:39:06ZengPublic Library of Science (PLoS)PLoS ONE1932-62032011-01-01612e2928910.1371/journal.pone.0029289A novel 'gene insertion/marker out' (GIMO) method for transgene expression and gene complementation in rodent malaria parasites.Jing-wen LinTakeshi AnnouraMohammed SajidSéverine Chevalley-MaurelJai RamesarOnny KlopBlandine M D Franke-FayardChris J JanseShahid M KhanResearch on the biology of malaria parasites has greatly benefited from the application of reverse genetic technologies, in particular through the analysis of gene deletion mutants and studies on transgenic parasites that express heterologous or mutated proteins. However, transfection in Plasmodium is limited by the paucity of drug-selectable markers that hampers subsequent genetic modification of the same mutant. We report the development of a novel 'gene insertion/marker out' (GIMO) method for two rodent malaria parasites, which uses negative selection to rapidly generate transgenic mutants ready for subsequent modifications. We have created reference mother lines for both P. berghei ANKA and P. yoelii 17XNL that serve as recipient parasites for GIMO-transfection. Compared to existing protocols GIMO-transfection greatly simplifies and speeds up the generation of mutants expressing heterologous proteins, free of drug-resistance genes, and requires far fewer laboratory animals. In addition we demonstrate that GIMO-transfection is also a simple and fast method for genetic complementation of mutants with a gene deletion or mutation. The implementation of GIMO-transfection procedures should greatly enhance Plasmodium reverse-genetic research.https://www.ncbi.nlm.nih.gov/pmc/articles/pmid/22216235/pdf/?tool=EBI |
| spellingShingle | Jing-wen Lin Takeshi Annoura Mohammed Sajid Séverine Chevalley-Maurel Jai Ramesar Onny Klop Blandine M D Franke-Fayard Chris J Janse Shahid M Khan A novel 'gene insertion/marker out' (GIMO) method for transgene expression and gene complementation in rodent malaria parasites. PLoS ONE |
| title | A novel 'gene insertion/marker out' (GIMO) method for transgene expression and gene complementation in rodent malaria parasites. |
| title_full | A novel 'gene insertion/marker out' (GIMO) method for transgene expression and gene complementation in rodent malaria parasites. |
| title_fullStr | A novel 'gene insertion/marker out' (GIMO) method for transgene expression and gene complementation in rodent malaria parasites. |
| title_full_unstemmed | A novel 'gene insertion/marker out' (GIMO) method for transgene expression and gene complementation in rodent malaria parasites. |
| title_short | A novel 'gene insertion/marker out' (GIMO) method for transgene expression and gene complementation in rodent malaria parasites. |
| title_sort | novel gene insertion marker out gimo method for transgene expression and gene complementation in rodent malaria parasites |
| url | https://www.ncbi.nlm.nih.gov/pmc/articles/pmid/22216235/pdf/?tool=EBI |
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