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In vivo trans-splicing of 5' and 3' segments of pre-mRNA directed by corresponding DNA sequences delivered by gene transfer.
| Publication Type | Academic Article |
| Authors | Pergolizzi R, Ropper A, Dragos R, Reid A, Nakayama K, Tan Y, Ehteshami J, Coleman S, Silver R, Hackett N, Menez A, Crystal R |
| Journal | Mol Ther |
| Volume | 8 |
| Issue | 6 |
| Pagination | 999-1008 |
| Date Published | 12/01/2003 |
| ISSN | 1525-0016 |
| Keywords | Gene Transfer Techniques, RNA Precursors, Trans-Splicing |
| Abstract | We have developed a new paradigm of in vivo gene transfer termed "segmental trans-splicing" (STS), in which individual "donor" and "acceptor" DNA sequences, delivered in vitro or in vivo, generate pre-mRNAs with 5' and 3' splice signals, respectively, and complementary hybridization domains through which the two pre-mRNAs interact, facilitating trans-splicing of the two mRNA fragments. To demonstrate STS, we used alpha-cobratoxin, a neurotoxin that binds irreversibly to postsynaptic nicotinic acetylcholine receptors. Cells or animals receiving both donor and acceptor plasmids, but neither plasmid alone, yielded RT-PCR products with the correct sequence of mature alpha-cobratoxin mRNA, suggesting that trans-splicing had occurred. Mice receiving intravenous administration of > or = 7.5 microg donor + acceptor plasmids, but not either plasmid alone, died within 6 h. These data demonstrate that segmental trans-splicing occurs in vivo. This approach should permit the intracellular assembly of molecules hitherto too large to be accommodated within current gene transfer vectors. |
| DOI | 10.1016/j.ymthe.2003.08.022 |
| PubMed ID | 14664803 |