Trypanosomes as a model to investigate mRNA decay pathways.
In trypanosomes, individual mRNAs arise by the processing of primary polycistronic transcripts. Consequently, mRNA degradation rates are critical determinants of mRNA abundance. In this chapter, we summarize the various options for genetic manipulation in trypanosomes with the goal of analyzing mRNA stability, including RNA interference. We describe a method for measuring the half-lives of trypanosome mRNAs, including those that are very unstable, and also the isolation of tagged protein-RNA complexes by IgG affinity chromatography. Last, we detail our current methods for RNA analysis with microarrays.
Archer
Stuart
S
Zentrum für Molekulare Biologie der Universität Heidelberg, Heidelberg, Germany.
Queiroz
Rafael
R
Stewart
Mhairi
M
Clayton
Christine
C
D000818:Animals; D015536:Down-Regulation; D006801:Humans; D007700:Kinetics; D008957:Models, Genetic; D020411:Oligonucleotide Array Sequence Analysis; D015800:Protozoan Proteins / Q000235:genetics / Q000378:metabolism; D020871:RNA Stability; D014345:Trypanosoma / Q000235:genetics*
D015800:Protozoan Proteins