Presence of an intron in inverted repeat constructs does not necessarily have an effect on efficiency of post-transcriptional gene silencing
- 81 Downloads
The effect of introns on silencing efficiency was tested in inverted repeat constructs of Granule-Bound Starch Synthase (GBSSI) cDNA by comparing the silencing efficiencies induced by inverted repeat constructs with and without introns. No effect could be attributed to the presence of introns indicating that the introns neither enhance nor inhibit post-transcriptional gene silencing. The effect of a spliceable intron in the spacer was studied by comparing constructs harbouring a spliceable or a non-spliceable intron in the spacer. As opposed to the general belief that splicing of an intron increases silencing efficiency, the use of a spliceable intron in the spacer did not result in enhancement of silencing in our experimental system.
Key words:Granule-bound starch synthase Intron Inverted repeat Post-transcriptional gene silencing Potato
Granule-Bound Starch Synthase I
Post-Transcriptional Gene Silencing
small interfering RNA
Unable to display preview. Download preview PDF.
This work was supported by the European Union (project QLK3-2000-00078). We would like to thank Mark Fiers for his help with the in silico predictons of the dsRNA structures, and Marcos Malosetti for statistical assistance.
- Heilersig H.J.B., Loonen A., Bergervoet M., Wolters A.M.A. and Visser R.G.F. 2006. Post-transcriptional gene silencing in potato: effects of size and sequence of the inverted repeats. Plant Mol. Biol. (in press).Google Scholar
- Hofvander P., Persson P.T., Tallberg P.T. and Wikstroem O. 1992. Genetically engineered modification of potato from amylopectin-type starch. International Patent Application WO 92/11376Google Scholar
- Sambrook J., Fritsch E.F., Maniatis T. (1989). Molecular Cloning: A Laboratory Manual, 2nd edn. Cold Spring Harbor, Cold Spring Harbor Laboratory PressGoogle Scholar
- Visser R.G.F., Hergersberg M., van der Leij F.R., Jacobsen E., Witholt B., Feenstra W.J. (1989). Molecular cloning and partial characterization of the gene for granule-bound starch synthase from a wild type and an amylose-free potato (Solanum tuberosum L.). Plant Sci. 64:185–192CrossRefGoogle Scholar
- Wesley S.V., Helliwell C.A., Smith N.A., Wang M.B., Rouse D.T., Liu Q., Gooding P.S., Singh S.P., Abbott D., Stoutjesdijk P.A., Robinson S.P., Gleave A.P., Green A.G., Waterhouse P.M. (2001). Construct design for efficient, effective and high-throughput gene silencing in plants. Plant J. 27:581–590CrossRefPubMedGoogle Scholar