Abstract
The Tetrahymena group I intron ribozyme is an RNA molecule of approx 400 bases that is capable of base-specific RNA trans-splicing. These properties can be applied to repair mutations at the RNA level, and therefore can restore normal cellular functions in diseased cells. The purpose of this chaper is to present the methodology that can be used to repair mutations responsible for myotonic dystrophy. The design and construction of group I intron ribozymes and their transfection in myotonic dystrophy cells will be among the sections described in this chapter.
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Cech, T. R., Zaug, A. J., and Grabowski, P. J. (1981) In vitro splicing of the ribosomal RNA precursor of Tetrahymena: involvement of a guanosine nucleotide in the excision of the intervening sequence. Cell 27, 487–496.
Sullenger, B. A. and Cech, T. R. (1994) Ribozyme-mediated repair of defective mRNA by targeted, trans-splicing. Nature 371, 619–622.
Phylactou, L. A., Darrah, C., and Wood, M. J. A. (1998) Ribozyme-mediated trans-splicing of a trinucleotide repeat. Nat. Genet. 18, 378–381.
Lan, N. Howery, R. P., Lee, S.-W., Smith, C. A., and Sullenger, B. A. (1998) Ribozyme-mediated repair of sickle beta-globin mRNAs in erythrocyte precursors. Science 280, 1593–1596.
Watanabe, T. and Sullenger, B. A. (2000) Induction of wild-type p53 activity in human cancer cells by ribozymes that repair mutant p53 transcripts. Proc. Natl. Acad. Sci. USA 97, 8490–8494.
Jones, J. T., Lee, S.-W., and Sullenger, B. A. (1996) Tagging ribozyme reaction sites to follow trans-splicing in mammalian cells. Nat. Med. 2, 643–648.
Kohler, U., Ayre, B. G., Goodman, H. M., and Haseloff, J. (1999) Trans-splicing ribozymes for targeted gene delivery. J. Mol. Biol. 285, 1935–1950.
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© 2004 Humana Press Inc., Totowa, NJ
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Phylactou, L.A. (2004). Repair of Myotonic Dystrophy Protein Kinase (DMPK) Transcripts by Trans-Splicing Ribozymes. In: Sioud, M. (eds) Ribozymes and siRNA Protocols. Methods in Molecular Biology™, vol 252. Humana Press. https://doi.org/10.1385/1-59259-746-7:373
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DOI: https://doi.org/10.1385/1-59259-746-7:373
Publisher Name: Humana Press
Print ISBN: 978-1-58829-226-1
Online ISBN: 978-1-59259-746-8
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