Analysis of gene function in Trypanosoma brucei using RNA interference.

Appolinaire Djikeng*, Shuiyuan Shen, Christian Tschudi, Elisabetta Ullu

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

Abstract / Description of output

Trypanosoma brucei, a flagellate protozoa of the family Trypanosomatidae, has become one of the model systems for unicellular pathogens to study fundamentally important biological phenomena. The method of choice today to examine gene function in these organisms is RNA interference (RNAi). Messenger RNA (mRNA) degradation is triggered by double-stranded RNA (dsRNA) produced in vivo from transgenes transcribed from opposing tetracycline (tet)-inducible T7 RNA polymerase promoters, or hairpin RNA transcribed from the tet-inducible procyclic acidic repetitive protein promoter. This chapter describes some of the methods we employ for ablation of gene expression by RNAi in T. brucei with particular emphasis on transfection and cloning of procyclic cells, induction of dsRNA expression, isolation of RNA, and analysis of dsRNA and target mRNA.

Original languageEnglish
Pages (from-to)287-298
Number of pages12
JournalMethods in molecular biology (Clifton, N.J.)
Publication statusPublished - 1 Jan 2004


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