Abstract |
Dystrophia myotonica type 1 (DM1), the most common muscular dystrophy in adults, results from expansion of a CTG repeat in the 3'-untranslated region of the dystrophia myotonica protein kinase gene (DMPK). Correction of the mutant DMPK transcript is a potential therapeutic strategy in DM1. We investigated the efficacy of artificial trans-splicing molecules (ATMs) to target and correct DMPK transcripts. ATMs designed to target intron 14 of DMPK pre-mRNA transcripts were tested for their ability to trans-splice the transcripts of a DMPK mini-gene construct and the endogenous DMPK transcripts of human myosarcoma cells (CCL-136). On agarose gel electrophoresis analysis, six of eight ATMs showed trans-splicing efficacy when applied to DMPK mini-gene construct transcripts, of which three were able to trans-splice endogenous DMPK pre-mRNA transcripts in myosarcoma cells, with trans-splicing efficiency ranging from 1.81 to 7.41%. These findings confirm that artificial trans-splicing can repair DMPK pre-mRNA and provide proof-of-principle evidence for this approach as potential therapeutic strategy for DM1.
|
Authors | H Y Chen, P Kathirvel, W C Yee, P S Lai |
Journal | Gene therapy
(Gene Ther)
Vol. 16
Issue 2
Pg. 211-7
(Feb 2009)
ISSN: 1476-5462 [Electronic] England |
PMID | 18923454
(Publication Type: Journal Article, Research Support, Non-U.S. Gov't)
|
Chemical References |
- DMPK protein, human
- RNA Precursors
- Myotonin-Protein Kinase
- Protein Serine-Threonine Kinases
|
Topics |
- Base Sequence
- Gene Targeting
(methods)
- Humans
- Molecular Sequence Data
- Myosarcoma
(enzymology, genetics)
- Myotonic Dystrophy
(enzymology, genetics)
- Myotonin-Protein Kinase
- Protein Serine-Threonine Kinases
(genetics)
- RNA Precursors
(genetics)
- Reverse Transcriptase Polymerase Chain Reaction
(methods)
- Trans-Splicing
(genetics)
- Transfection
- Tumor Cells, Cultured
|