Abstract
In 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.
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Acknowledgements
We thank all members of the group headed by M Bach-Elias for their comments on the manuscript. This work was supported by the Asociación Española contra el Cáncer, La Marató de TV3 and Fundación Ramón Areces. S Guil was a recipient of a BEFI fellowship. We also thank Martí Cullell for revising this manuscript. We thank Dr AD Levinson, Dr Y Shimura and Dr I Mattaj for donating the c-H-ras genes, the ASLV ESE sequence and the adenovirus sequence, respectively.
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Codony, C., Guil, S., Caudevilla, C. et al. Modulation in vitro of H-ras oncogene expression by trans-splicing. Oncogene 20, 3683–3694 (2001). https://doi.org/10.1038/sj.onc.1204473
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DOI: https://doi.org/10.1038/sj.onc.1204473