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

Area of Science:

• Molecular Biology
• Parasitology
• Gene Regulation

Background:

• Leishmania parasites present unique challenges for genetic manipulation.
• Controlled gene expression is crucial for understanding parasite biology and developing interventions.

Purpose of the Study:

• To establish a novel inducible gene expression system in Leishmania.
• To validate the system's efficacy and applicability for gene function analysis.

Main Methods:

• Development of a dimerizable Cre recombinase (DiCre) system.
• Induction of gene expression using rapamycin.
• Analysis of gene product expression in integrated and episomal contexts.
• Validation using the checkpoint protein Rad9.

Main Results:

• Rapamycin-induced DiCre activation enabled efficient, time- and dose-dependent gene flipping and expression.
• The system functions effectively in both integrated and episomal genetic contexts.
• Expression of Rad9 variants revealed the C-terminal domain's importance for subcellular localization.

Conclusions:

• The DiCre-based inducible system is a robust tool for controlled gene expression in Leishmania.
• This system significantly advances the study of gene function in this important protozoan parasite.
• The findings provide new insights into the function of the Rad9 protein in Leishmania.