In vivo analysis of the RNA interference mechanism in Trypanosoma brucei

Christian Tschudi, Appolinaire Djikeng, Huafang Shi, Elisabetta Ullu*

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

Abstract

Flagellate protozoa of the family Trypanosomatidae, which includes various members of the genera Leishmania and Trypanosoma, are model systems for unicellular pathogens to study fundamentally important biological phenomena. Recently, ablation of gene expression by RNA interference (RNAi) has become the method of choice to study gene function in Trypanosoma brucei, an early divergent eukaryote that infects humans and animals. As has been shown in multicellular organisms, the RNAi mechanism in T. brucei involves processing of double-stranded RNA 24- to 26-nt RNAs, termed small interfering RNAs (siRNAs), which guide degradation of the target mRNA. In this article, we describe some of the methods we employ for the analysis of the RNAi mechanism in T. brucei with particular emphasis on detection, cloning, and fractionation of siRNAs and siRNA complexes.

Original languageEnglish
Pages (from-to)304-312
Number of pages9
JournalMethods
Volume30
Issue number4
DOIs
Publication statusPublished - 1 Aug 2003

Keywords

  • Cloning of siRNAs
  • Detection of siRNAs
  • Double-stranded RNA
  • Inducible RNAi
  • Polysome gradient
  • RNA interference
  • Trypanosome

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