Single-locus targeting constructs for reliable regulated RNAi and transgene expression in Trypanosoma brucei

Molecular and Biochemical Parasitology

Volumn 161, Issue 1, 2008, Pages 76-79

  Alsford, Sam ^a   Horn, David ^a  

^a LONDON SCHOOL OF HYGIENE AND TROPICAL MEDICINE   (United Kingdom)

Author keywords

Epigenetic; Functional genomics; Localisation; Position effects; RNAi

Indexed keywords

RECOMBINANT PROTEIN;

ARTICLE; BLOOD FLOW; CELLULAR DISTRIBUTION; CHROMATIN; EPIGENETICS; FUNCTIONAL GENOMICS; GENE CONSTRUCT; GENE EXPRESSION; GENE EXPRESSION REGULATION; GENE LOCUS; GENE TARGETING; GENETIC TRANSCRIPTION; NONHUMAN; ONCOGENE C MYC; POSITION EFFECT; PRIORITY JOURNAL; PROTEIN INTERACTION; RELIABILITY; REPRODUCIBILITY; RNA INTERFERENCE; TRANSGENE; TRYPANOSOMA BRUCEI;

ANIMALS; GENE EXPRESSION; GENE SILENCING; GENE TARGETING; RECOMBINANT PROTEINS; RNA, DOUBLE-STRANDED; TRANSGENES; TRYPANOSOMA BRUCEI BRUCEI;

TRYPANOSOMA BRUCEI;

EID: 47949091926     PISSN: 01666851     EISSN: None     Source Type: Journal    
DOI: 10.1016/j.molbiopara.2008.05.006     Document Type: Article

References (20)

1. Berriman M., Ghedin E., Hertz-Fowler C., et al. The genome of the African trypanosome Trypanosoma brucei. Science 309 (2005) 416-422
2. Kelly S., Reed J., Kramer S., et al. Functional genomics in Trypanosoma brucei: a collection of vectors for the expression of tagged proteins from endogenous and ectopic gene loci. Mol Biochem Parasitol 154 (2007) 103-109
3. Oberholzer M., Morand S., Kunz S., and Seebeck T. A vector series for rapid PCR-mediated C-terminal in situ tagging of Trypanosoma brucei genes. Mol Biochem Parasitol 145 (2006) 117-120
4. Shen S., Arhin G.K., Ullu E., and Tschudi C. In vivo epitope tagging of Trypanosoma brucei genes using a one step PCR-based strategy. Mol Biochem Parasitol 113 (2001) 171-173
5. Alibu V.P., Storm L., Haile S., Clayton C., and Horn D. A doubly inducible system for RNA interference and rapid RNAi plasmid construction in Trypanosoma brucei. Mol Biochem Parasitol 139 (2005) 75-82
6. LaCount D.J., Bruse S., Hill K.L., and Donelson J.E. Double-stranded RNA interference in Trypanosoma brucei using head-to-head promoters. Mol Biochem Parasitol 111 (2000) 67-76
7. Morris J.C., Wang Z., Drew M.E., Paul K.S., and Englund P.T. Inhibition of bloodstream form Trypanosoma brucei gene expression by RNA interference using the pZJM dual T7 vector. Mol Biochem Parasitol 117 (2001) 111-113
8. Wirtz E., and Clayton C. Inducible gene expression in trypanosomes mediated by a prokaryotic repressor. Science 268 (1995) 1179-1183
9. Das A., Banday M., and Bellofatto V. RNA polymerase transcription machinery in trypanosomes. Eukaryot Cell 7 (2008) 429-434
10. Wirtz E., Leal S., Ochatt C., and Cross G.A. A tightly regulated inducible expression system for conditional gene knock-outs and dominant-negative genetics in Trypanosoma brucei. Mol Biochem Parasitol 99 (1999) 89-101
11. Alsford S., Kawahara T., Glover L., and Horn D. Tagging a T. brucei RRNA locus improves stable transfection efficiency and circumvents inducible expression position effects. Mol Biochem Parasitol 144 (2005) 142-148
12. Glover L., Alsford S., Beattie C., and Horn D. Deletion of a trypanosome telomere leads to loss of silencing and progressive loss of terminal DNA in the absence of cell cycle arrest. Nucleic Acids Res 35 (2007) 872-880
13. White T.C., Rudenko G., and Borst P. Three small RNAs within the 10 kb trypanosome rRNA transcription unit are analogous to domain VII of other eukaryotic 28S rRNAs. Nucleic Acids Res 14 (1986) 9471-9489
14. Markstein M., Pitsouli C., Villalta C., Celniker S.E., and Perrimon N. Exploiting position effects and the gypsy retrovirus insulator to engineer precisely expressed transgenes. Nat Genet 40 (2008) 476-483
15. Motyka S.A., Zhao Z., Gull K., and Englund P.T. Integration of pZJM library plasmids into unexpected locations in the Trypanosoma brucei genome. Mol Biochem Parasitol 134 (2004) 163-167
16. Alsford S., and Horn D. RNA polymerase I transcription stimulates homologous recombination in Trypanosoma brucei. Mol Biochem Parasitol 153 (2007) 77-79
17. Subramaniam C., Veazey P., Redmond S., et al. Chromosome-wide analysis of gene function by RNA interference in the African trypanosome. Eukaryot Cell 5 (2006) 1539-1549
18. Durand-Dubief M., Kohl L., and Bastin P. Efficiency and specificity of RNA interference generated by intra- and intermolecular double stranded RNA in Trypanosoma brucei. Mol Biochem Parasitol 129 (2003) 11-21
19. Horn D. Codon usage suggests that translational selection has a major impact on protein expression in trypanosomatids. BMC Genom 9 (2008) 2
20. Redmond S., Vadivelu J., and Field M.C. RNAit: an automated web-based tool for the selection of RNAi targets in Trypanosoma brucei. Mol Biochem Parasitol 128 (2003) 115-118