Development of a toolkit for piggyBac-mediated integrative transfection of the human filarial parasite Brugia malayi

PLoS Neglected Tropical Diseases
Canhui LiuThomas R Unnasch

Abstract

The human filarial parasites cause diseases that are among the most important causes of morbidity in the developing world. The elimination programs targeting these infections rely on a limited number of drugs, making the identification of new chemotherapeutic agents a high priority. The study of these parasites has lagged due to the lack of reverse genetic methods. We report a novel co-culture method that results in developmentally competent infective larvae of one of the human filarial parasites (Brugia malayi) and describe a method to efficiently transfect the larval stages of this parasite. We describe the production of constructs that result in integrative transfection using the piggyBac transposon system, and a selectable marker that can be used to identify transgenic parasites. We describe the production and use of dual reporter plasmids containing both a secreted luciferase selectable marker and fluorescent protein reporters that will be useful to study temporal and spatial patterns of gene expression. The methods and constructs reported here will permit the efficient production of integrated transgenic filarial parasite lines, allowing reverse genetic technologies to be applied to all life cycle stages of the parasite.

References

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Citations

Apr 4, 2020·PLoS Neglected Tropical Diseases·Canhui LiuThomas R Unnasch
Jun 24, 2020·PLoS Neglected Tropical Diseases·Alexandra GroteElodie Ghedin
Sep 1, 2020·PLoS Neglected Tropical Diseases·Canhui LiuThomas R Unnasch
Feb 9, 2020·The Journal of Experimental Biology·Michelle L CastellettoElissa A Hallem
Feb 14, 2021·Biomedicine & Pharmacotherapy = Biomédecine & Pharmacothérapie·Alexander KwartengSamuel Terkper Ahuno

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Datasets Mentioned

BETA
HQ388295

Methods Mentioned

BETA
transfection
transgenic
PCR
lavage
electrophoresis
transfections

Software Mentioned

AxioVision

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