Inducible expression of suicide genes in Leishmania donovani amastigotes.
This study tests the feasibility of using the A2 gene regulatory system to create a Leishmania cell line in which attenuation is developmentally regulated when the parasite differentiates from promastigotes to amastigotes. The Leishmania donovani- inducible A2 gene regulatory system was used to differentially express in amastigotes two potential suicide genes: a truncated version of the L. donovani 3'-nucleotidase/nuclease expressed in the cytoplasm and the herpes simplex virus thymidine kinase gene. These genes were inserted between A2 noncoding regulatory sequences for up-regulation of expression in amastigotes. The accumulation of toxic products affected L. donovani cell replication and viability both in vitro and in vivo. The inducible expression of toxic gene products represents a valuable tool for the development of safe and effective vaccines.
Institute of Parasitology, McGill University, Macdonald Campus, Ste-Anne de Bellevue, Quebec H9X 3V9, Canada.
E
E
Ghedin
H
H
Charest
W W
WW
Zhang
A
A
Debrabant
D
D
Dwyer
G
G
Matlashewski
D000818:Animals; D002454:Cell Differentiation; D004351:Drug Resistance / Q000235:genetics; D005786:Gene Expression Regulation; D018390:Gene Targeting; D017125:Genes, Protozoan; D007893:Leishmania donovani / Q000166:cytology / Q000235:genetics*; D009355:Neomycin / Q000494:pharmacology; D009708:Nucleotidases / Q000235:genetics; D016052:Protozoan Vaccines / Q000235:genetics; D012045:Regulatory Sequences, Nucleic Acid; D013937:Thymidine Kinase / Q000235:genetics; D014162:Transfection; D014613:Vaccines, Attenuated / Q000235:genetics
D016052:Protozoan Vaccines; D014613:Vaccines, Attenuated; D013937:Thymidine Kinase; D009708:Nucleotidases; C040470:3'-nucleotidase; D009355:Neomycin