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Transient transfection and expression of foreign and endogenous genes in the intracellular stages of Trypanosoma cruzi.

Abstract：

:The capacity for rapid localization of epitope-tagged or fluorescent fusion proteins in cells is an important tool for biological discovery and functional analysis. For Trypanosoma cruzi, the protozoan parasite that causes human Chagas disease, visualization of ectopically-expressed proteins in the clinically-relevant mammalian stages is hindered by the necessity to first perform transfection and lengthy selection procedures in the insect vector form of the parasite. Here, we demonstrate the ability to by-pass the insect stage with the delivery of plasmid DNA to non-dividing, tissue culture trypomastigotes such that upon host cell infection, transgenes are expressed and rapidly localized in intracellular T. cruzi amastigotes. The inclusion of a sorting step prior to host cell infection by trypomastigotes greatly enriches (>90%) the number of transgene-expressing amastigotes observed in mammalian host cells. This is a significant methodological advance that has the potential to accelerate the pace of discovery in the Chagas disease field.

journal_name

Mol Biochem Parasitol

journal_title

Molecular and biochemical parasitology

authors

Padmanabhan PK,Polidoro RB,Barteneva NS,Gazzinelli RT,Burleigh BA

doi

10.1016/j.molbiopara.2015.02.001

subject

Has Abstract

pub_date

2014-12-01 00:00:00

pages

100-3

issue

2

eissn

0166-6851

issn

1872-9428

pii

S0166-6851(15)00007-9

journal_volume

198

pub_type

杂志文章

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