Modulating alternative splicing by cotranscriptional cleavage of nascent intronic RNA
- 1Sir William Dunn School of Pathology, University of Oxford, Oxford, OX1 3RE, United Kingdom
- 2International Centre for Genetic Engineering and Biotechnology, 34012 Trieste, Italy
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↵3 These authors contributed equally to this work.
Abstract
Cotranscriptional cleavage mediated by a hammerhead ribozyme can affect alternative splicing if interposed between an exon and its intronic regulatory elements. This has been demonstrated using two different alternative splicing systems based on α-tropomyosin and fibronectin genes. We suggest that there is a requirement for intronic regulatory elements to be covalently attached to exons that are in turn tethered to the elongating polymerase. In the case of the alternatively spliced EDA exon of the fibronectin gene, we demonstrate that the newly identified intronic downstream regulatory element is associated with the splicing regulatory protein SRp20. Our results suggest that targeted hammerhead ribozyme cleavage within introns can be used as a tool to define splicing regulatory elements.
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Footnotes
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Reprint requests to: Nicholas J. Proudfoot, Sir William Dunn School of Pathology, University of Oxford, Oxford, OX1 3RE, United Kingdom; e-mail: nicholas.proudfoot{at}path.ox.ac.uk; fax 86-5-275556; or Franco Pagani, International Centre for Genetic Engineering and Biotechnology, Padriciano 99, 34012 Trieste, Italy; e-mail: pagani{at}icgeb.org; fax 86-5-275556.
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Article published online ahead of print. Article and publication date are at http://www.rnajournal.org/cgi/doi/10.1261/rna.615508.
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- Received May 2, 2007.
- Accepted October 24, 2007.
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Freely available online through the open access option.
- Copyright © 2008 RNA Society
