A DiCre recombinase-based system for inducible expression in Leishmania major

Renato E R S Santos; Gabriel L A Silva; Elaine V Santos; Samuel M Duncan; Jeremy C Mottram; Jeziel D Damasceno; Luiz R O Tosi

doi:10.1016/j.molbiopara.2017.06.006

A DiCre recombinase-based system for inducible expression in Leishmania major

Renato E R S Santos, Gabriel L A Silva, Elaine V Santos, Samuel M Duncan, Jeremy C Mottram, Jeziel D Damasceno, Luiz R O Tosi

Biology

Research output: Contribution to journal › Article › peer-review

Abstract

Here we present the establishment of an inducible system based on the dimerizable Cre recombinase (DiCre) for controlled gene expression in the protozoan parasite Leishmania. Rapamycin-induced DiCre activation promoted efficient flipping and expression of gene products in a time and dose-dependent manner. The DiCre flipping activity induced the expression of target genes from both integrated and episomal contexts broadening the applicability of the system. We validated the system by inducing the expression of both full length and truncated forms of the checkpoint protein Rad9, which revealed that the highly divergent C-terminal domain of Rad9 is necessary for proper subcellular localization. Thus, by establishing the DiCre-based inducible system we have created and validated a robust new tool for assessing gene function in Leishmania.

Original language	English
Pages (from-to)	45-48
Number of pages	4
Journal	MOLECULAR AND BIOCHEMICAL PARASITOLOGY
Volume	216
Early online date	16 Jun 2017
DOIs	https://doi.org/10.1016/j.molbiopara.2017.06.006
Publication status	Published - 1 Sept 2017

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Journal Article

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10.1016/j.molbiopara.2017.06.006

SANTOS et al MS
© 2017 Elsevier B.V. All rights reserved. This is an author-produced version of the published paper. Uploaded in accordance with the publisher’s self-archiving policy.
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Cite this

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abstract = "Here we present the establishment of an inducible system based on the dimerizable Cre recombinase (DiCre) for controlled gene expression in the protozoan parasite Leishmania. Rapamycin-induced DiCre activation promoted efficient flipping and expression of gene products in a time and dose-dependent manner. The DiCre flipping activity induced the expression of target genes from both integrated and episomal contexts broadening the applicability of the system. We validated the system by inducing the expression of both full length and truncated forms of the checkpoint protein Rad9, which revealed that the highly divergent C-terminal domain of Rad9 is necessary for proper subcellular localization. Thus, by establishing the DiCre-based inducible system we have created and validated a robust new tool for assessing gene function in Leishmania.",

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AU - Santos, Renato E R S

AU - Silva, Gabriel L A

AU - Santos, Elaine V

AU - Duncan, Samuel M

AU - Mottram, Jeremy C

AU - Damasceno, Jeziel D

AU - Tosi, Luiz R O

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