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

Mol Biochem Parasitol. 2017 Sep:216:45-48. doi: 10.1016/j.molbiopara.2017.06.006. Epub 2017 Jun 16.

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.

Keywords: 9-1-1 complex; DNA damage response; DiCre recombinase; Inducible expression; Leishmania; Rad9-Rad1-Hus1.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Cell Cycle Proteins / metabolism
  • Gene Expression Regulation*
  • Gene Order
  • Genetic Engineering*
  • Genetic Vectors / genetics
  • Homologous Recombination*
  • Integrases / metabolism*
  • Leishmania major / genetics*
  • Leishmania major / metabolism*

Substances

  • Cell Cycle Proteins
  • rad9 protein
  • Cre recombinase
  • Integrases