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...

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Autores principales:	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
Formato:	article
Lenguaje:	EN
Publicado:	Public Library of Science (PLoS) 2011
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/9f23ec21c36444b8b6062e927838ab68
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spelling	oai:doaj.org-article:9f23ec21c36444b8b6062e927838ab682021-11-18T07:31:31ZA novel 'gene insertion/marker out' (GIMO) method for transgene expression and gene complementation in rodent malaria parasites.1932-620310.1371/journal.pone.0029289https://doaj.org/article/9f23ec21c36444b8b6062e927838ab682011-01-01T00:00:00Zhttps://www.ncbi.nlm.nih.gov/pmc/articles/pmid/22216235/pdf/?tool=EBIhttps://doaj.org/toc/1932-6203Research 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.Jing-wen LinTakeshi AnnouraMohammed SajidSéverine Chevalley-MaurelJai RamesarOnny KlopBlandine M D Franke-FayardChris J JanseShahid M KhanPublic Library of Science (PLoS)articleMedicineRScienceQENPLoS ONE, Vol 6, Iss 12, p e29289 (2011)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q 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.
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.
format	article
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
title	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_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_sort	novel 'gene insertion/marker out' (gimo) method for transgene expression and gene complementation in rodent malaria parasites.
publisher	Public Library of Science (PLoS)
publishDate	2011
url	https://doaj.org/article/9f23ec21c36444b8b6062e927838ab68
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A novel 'gene insertion/marker out' (GIMO) method for transgene expression and gene complementation in rodent malaria parasites.

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