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Modulation in vitro of H-ras oncogene expression by trans-splicing

AuthorsCodony, Carles; Guil, Sonia ; Caudevilla, Concha; Serra, Dolors; Asins, Guillermina; Graessmann, Adolf; Hegardt, Fausto G.; Bach-Elias, Montse
Issue Date21-Jun-2001
PublisherNature Publishing Group
CitationOncogene 20(28): 3683-3694 (2001)
AbstractIn man, activated N-, K- and H-ras oncogenes have been found in around 30% of the solid tumours tested. An exon known as IDX, which has been described previously and is located between exon 3 and exon 4A of the c-H-ras pre-mRNA, allows an alternative splicing process that results in the synthesis of the mRNA of a putative protein named p19. It has been suggested that this alternative pathway is less tumorigenic than that which results in the activation of p21. We have used the mammalian trans-splicing mechanism as a tool with which to modulate this particular pre-mRNA processing to produce mRNA similar to that of mature p19 RNA. The E4A exon of the activated H-ras gene was found to be a good target for external trans-splicing. We reprogrammed the rat carnitine octanoyltransferase exon 2 to specifically invade the terminal region of H-ras. Assays performed with this reprogrammed trans-exon showed that the trans-splicing product was obtained in competition with cis-splicing of the D intron of the H-ras gene, and was associated with concomitant down-modulation of D intron cis-splicing. We also found that the exon 4A of the human c-H-ras gene underwent successive trans-splicing rounds with an external exon.
Publisher version (URL)http://dx.doi.org/10.1038/sj.onc.1204473
Identifiersdoi: 10.1038/sj.onc.1204473
issn: 0950-9232
e-issn: 1476-5594
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