Splicing of mRNA mediated by tRNA sequences in mouse cells
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↵1 These authors contributed equally to this work.
Abstract
tRNA splicing is essential for the formation of tRNAs and therefore for gene expression. A circularly permuted sequence of an amber-suppressor pre-tRNA gene was inserted into the sequence encoding the mouse NEMO protein. We demonstrated that, in mouse cells, the hybrid pre-tRNA/pre-mRNAs can be spliced precisely at the sites of the pre-tRNA intron. This splicing reaction produces functional tRNAs that suppress amber codons as well as translatable mRNAs that sustain the NF-κB activation pathway. The RNA molecules extracted from mouse cells were amplified by RT-PCR, and their sequences were determined, confirming the identity of the splice junctions. We then applied the Archaea-express technology, in which an archaeal RNA endonuclease is expressed in mouse cells. We show that both the endogenous eukaryal endonuclease and the archaeal one cleave the hybrid pre-tRNA/pre-mRNAs in the same manner with an additive effect.
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Footnotes
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Reprint requests to: Glauco P. Tocchini-Valentini, Istituto di Biologia Cellulare, Consiglio Nazionale delle Ricerche, Via Ramarini 32, 00015 Monterotondo, Rome, Italy; e-mail: gtocchini{at}ibc.cnr.it; fax: 39-06-90091261.
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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.1841609.
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- Received July 22, 2009.
- Accepted August 27, 2009.
- Copyright © 2009 RNA Society