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Presence of an intron in inverted repeat constructs does not necessarily have an effect on efficiency of post-transcriptional gene silencing

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Abstract

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.

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Abbreviations

dsRNA:

double-stranded RNA

GBSSI:

Granule-Bound Starch Synthase I

IME:

intron-mediated enhancement

PTGS:

Post-Transcriptional Gene Silencing

siRNA:

small interfering RNA

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Acknowledgements

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.

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Authors and Affiliations

  1. Graduate School Experimental Plant Sciences, Laboratory of Plant Breeding, Department of Plant Sciences, Wageningen University, P.O. Box 386, 6700, AJ, Wageningen, the Netherlands

    Berlinda H.J.B. Heilersig, Annelies E.H.M. Loonen, Anne-Marie A. Wolters & Richard G.F. Visser

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  1. Berlinda H.J.B. Heilersig
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  2. Annelies E.H.M. Loonen
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  3. Anne-Marie A. Wolters
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  4. Richard G.F. Visser
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Correspondence to Anne-Marie A. Wolters.

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Heilersig, B., Loonen, A., Wolters, AM. et al. Presence of an intron in inverted repeat constructs does not necessarily have an effect on efficiency of post-transcriptional gene silencing. Mol Breeding 17, 307–316 (2006). https://doi.org/10.1007/s11032-006-9001-x

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  • DOI: https://doi.org/10.1007/s11032-006-9001-x

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