High efficacy in vitro selection procedure for generating transgenic parasites of Plasmodium berghei using an antibiotic toxic to rodent hosts

Abstract The malaria parasite Plasmodium berghei is one of the main rodent malaria models. A shortcoming of this model parasite is its low flexibility in genetic manipulation. As this parasite cannot be continuously propagated in cell cultures, in vivo drug selection procedures are necessary to isol...

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Autores principales:	Akira Soga, Hironori Bando, Mami Ko-ketsu, Hirono Masuda-Suganuma, Shin-ichiro Kawazu, Shinya Fukumoto
Formato:	article
Lenguaje:	EN
Publicado:	Nature Portfolio 2017
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/e952a447d54247978f819451a93f4600
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spelling	oai:doaj.org-article:e952a447d54247978f819451a93f46002021-12-02T12:30:44ZHigh efficacy in vitro selection procedure for generating transgenic parasites of Plasmodium berghei using an antibiotic toxic to rodent hosts10.1038/s41598-017-04244-02045-2322https://doaj.org/article/e952a447d54247978f819451a93f46002017-06-01T00:00:00Zhttps://doi.org/10.1038/s41598-017-04244-0https://doaj.org/toc/2045-2322Abstract The malaria parasite Plasmodium berghei is one of the main rodent malaria models. A shortcoming of this model parasite is its low flexibility in genetic manipulation. As this parasite cannot be continuously propagated in cell cultures, in vivo drug selection procedures are necessary to isolate genetic mutants. Drugs harmful to rodents therefore cannot be used for drug selection, which restricts the range of genetic manipulation. In this study, we addressed this problem by establishing a novel in vitro culture drug selection method, which we used in combination with other established methods to successfully isolate genetically manipulated parasites. The target mutants were enriched to the desired level within two weeks. We show that our system can also be used for sequential genetic manipulation of parasites carrying the traditionally used selection markers, demonstrate the procedure’s versatility, and show its use in isolating specific genetically manipulated parasites. This novel in vitro selection method increases the number of available selection markers, allowing more extensive genetic manipulation in malaria parasite research.Akira SogaHironori BandoMami Ko-ketsuHirono Masuda-SuganumaShin-ichiro KawazuShinya FukumotoNature PortfolioarticleMedicineRScienceQENScientific Reports, Vol 7, Iss 1, Pp 1-11 (2017)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Akira Soga Hironori Bando Mami Ko-ketsu Hirono Masuda-Suganuma Shin-ichiro Kawazu Shinya Fukumoto High efficacy in vitro selection procedure for generating transgenic parasites of Plasmodium berghei using an antibiotic toxic to rodent hosts
description	Abstract The malaria parasite Plasmodium berghei is one of the main rodent malaria models. A shortcoming of this model parasite is its low flexibility in genetic manipulation. As this parasite cannot be continuously propagated in cell cultures, in vivo drug selection procedures are necessary to isolate genetic mutants. Drugs harmful to rodents therefore cannot be used for drug selection, which restricts the range of genetic manipulation. In this study, we addressed this problem by establishing a novel in vitro culture drug selection method, which we used in combination with other established methods to successfully isolate genetically manipulated parasites. The target mutants were enriched to the desired level within two weeks. We show that our system can also be used for sequential genetic manipulation of parasites carrying the traditionally used selection markers, demonstrate the procedure’s versatility, and show its use in isolating specific genetically manipulated parasites. This novel in vitro selection method increases the number of available selection markers, allowing more extensive genetic manipulation in malaria parasite research.
format	article
author	Akira Soga Hironori Bando Mami Ko-ketsu Hirono Masuda-Suganuma Shin-ichiro Kawazu Shinya Fukumoto
author_facet	Akira Soga Hironori Bando Mami Ko-ketsu Hirono Masuda-Suganuma Shin-ichiro Kawazu Shinya Fukumoto
author_sort	Akira Soga
title	High efficacy in vitro selection procedure for generating transgenic parasites of Plasmodium berghei using an antibiotic toxic to rodent hosts
title_short	High efficacy in vitro selection procedure for generating transgenic parasites of Plasmodium berghei using an antibiotic toxic to rodent hosts
title_full	High efficacy in vitro selection procedure for generating transgenic parasites of Plasmodium berghei using an antibiotic toxic to rodent hosts
title_fullStr	High efficacy in vitro selection procedure for generating transgenic parasites of Plasmodium berghei using an antibiotic toxic to rodent hosts
title_full_unstemmed	High efficacy in vitro selection procedure for generating transgenic parasites of Plasmodium berghei using an antibiotic toxic to rodent hosts
title_sort	high efficacy in vitro selection procedure for generating transgenic parasites of plasmodium berghei using an antibiotic toxic to rodent hosts
publisher	Nature Portfolio
publishDate	2017
url	https://doaj.org/article/e952a447d54247978f819451a93f4600
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High efficacy in vitro selection procedure for generating transgenic parasites of Plasmodium berghei using an antibiotic toxic to rodent hosts

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