Development of a toolkit for piggyBac-mediated integrative transfection of the human filarial parasite Brugia malayi.
PLoS Negl Trop Dis
; 12(5): e0006509, 2018 05.
Article
en En
| MEDLINE
| ID: mdl-29782496
ABSTRACT
BACKGROUND:
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. METHODOLOGY/PRINCIPALFINDINGS:
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. CONCLUSIONS/SIGNIFICANCE:
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.
Texto completo:
1
Base de datos:
MEDLINE
Asunto principal:
Elementos Transponibles de ADN
/
Transfección
/
Brugia Malayi
Tipo de estudio:
Evaluation_studies
/
Prognostic_studies
Idioma:
En
Revista:
PLoS Negl Trop Dis
Asunto de la revista:
MEDICINA TROPICAL
Año:
2018
Tipo del documento:
Article