Development of an inducible protein expression system based on the protozoan host Leishmania tarentolae

The genetic toolbox forLeishmaniaparasites

“…53,54 A novel system utilizing artificial linear episome-based structures shows promise for expressing large quantities of target proteins in L. tarentolae for the purpose of protein purification. 55,56 A unique challenge to the development of regulated expression systems is posed by the unusual lack of transcriptional control in Leishmania parasites. Attempts to develop systems where gene expression can be induced have been disappointing thus far but are being explored further.…”

“…Attempts to develop systems where gene expression can be induced have been disappointing thus far but are being explored further. [56][57][58] As mentioned above, RNAi for knock-down of gene expression is not possible in most Leishmania species. However, a technique that regulates the stability of specific proteins has recently been modified for the use in Leishmania.…”

T7 polymerase-driven transcription is downregulated in metacyclic promastigotes and amastigotes of Leishmania mexicana

“…In our study, the mCherry, T7 polymerase, and Tet-repressor genes were flanked by the UTRs derived from the calmodulin (LtaP.09.0940) intergenic regions of Leishmania tarentolae Wenyon, 1921(Breitling et al 2002, Kushnir et al 2005. Whole-transcriptome profiling revealed that calmodulin mRNA abundance in species of Leishmania is developmentally regulated.…”

“…The construct was integrated into the same 18S rRNA locus (Kushnir et al 2005) and expression of the mCherry protein was confirmed by fluorescence microscopy (data not shown). The differentiation of the transgenic culture and proper separation of the life cycle stages were verified by (Biagi, 1953).…”

RNA polymerase I promoter and splice acceptor site recognition affect gene expression in non-pathogenic Leishmania species

“…It is also a useful alternative organism for the expression of accurately-modified heterologous eukaryotic proteins. A short doubling-time and simple nutrient requirements (Simpson & Braly 1970) make L. tarentolae an attractive host for high level production of heterologous proteins containing N-linked oligosaccharide modifications similar to those found in mammals (Breitling et al 2002, Kushnir et al 2005, and for generating isotopically-labeled protein for nuclear magnetic resonance studies (Niculae et al 2006). In addition, L. tarentolae can be used as a live vaccine capable of eliciting protective immune response against L. (Leishmania) donovani, opening a new avenue for vaccination against other species involved in cutaneous leishmaniasis without the risk of disease development in humans (Breton et al 2005).…”