Abstract
Induction of gene expression is a valuable approach for functional studies since it allows for the assessment of phenotypes without the need for clonal selection. Inducible expression can find a wide range of applications, from the study of essential genes to the characterization of overexpression of genes of interest. Here, we describe a detailed protocol for the use of the DiCre-based inducible gene expression system in Leishmania parasites. This is a tightly regulated induction system that allows for time- and dose-controlled expression of gene products, as rapidly as within 12 h.
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References
Santos RERS, Silva GLA, Santos EV et al (2017) A DiCre recombinase-based system for inducible expression in Leishmania major. Mol Biochem Parasitol 216:45–48. https://doi.org/10.1016/j.molbiopara.2017.06.006
Jullien N, Sampieri F, Enjalbert A, Herman J-P (2003) Regulation of Cre recombinase by ligand-induced complementation of inactive fragments. Nucleic Acids Res 31:e131. https://doi.org/10.1093/NAR/GNG131
Albert H, Dale EC, Lee E, Ow DW (1995) Site-specific integration of DNA into wild-type and mutant lox sites placed in the plant genome. Plant J 7:649–659. https://doi.org/10.1046/j.1365-313X.1995.704649.x
Kapler GM, Coburn CM, Beverley SM (1990) Stable transfection of the human parasite Leishmania major delineates a 30-kilobase region sufficient for extrachromosomal replication and expression. Mol Cell Biol 10:1084–1094. https://doi.org/10.1128/MCB.10.3.1084
Duncan SM, Myburgh E, Philipon C et al (2016) Conditional gene deletion with DiCre demonstrates an essential role for CRK3 in L. eishmania mexicana cell cycle regulation. Mol Microbiol 100:931–944. https://doi.org/10.1111/mmi.13375
Robinson KA, Beverley SM (2003) Improvements in transfection efficiency and tests of RNA interference (RNAi) approaches in the protozoan parasite Leishmania. Mol Biochem Parasitol 128:217–228. https://doi.org/10.1016/S0166-6851(03)00079-3
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Damasceno, J.D., Tosi, L.R.O., Santos, R.E.R.S., Mottram, J.C. (2019). DiCre-Based Inducible Gene Expression. In: Clos, J. (eds) Leishmania. Methods in Molecular Biology, vol 1971. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-9210-2_11
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DOI: https://doi.org/10.1007/978-1-4939-9210-2_11
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