Back to Search
Start Over
Skipping of Duplicated Dystrophin Exons: In Vitro Induction and Assessment
- Source :
- Methods in Molecular Biology ISBN: 9781493986507
- Publication Year :
- 2018
- Publisher :
- Springer New York, 2018.
-
Abstract
- Duplications of one or more dystrophin exons that disrupt the reading frame account for about 15% of all Duchenne cases, and like the more common genomic deletions, most pathogenic duplications of single or multiple dystrophin exons are also amenable to targeted exon skipping. However, additional considerations must be taken into account: (1) skipping of all duplicated exons, and, flanking exons as necessary, will frequently be required to restore the reading frame and generate an in-frame Becker muscular dystrophy-like mRNA, (2) the phosphorodiamidate morpholino oligomer chemistry is more effective than the 2'-O-methyl modified oligonucleotides at inducing multiple exon skipping, and (2) the apparent efficiency of exon skipping can be confounded by the choice of RT-PCR system. Standard RT-PCR systems can preferentially amplify the shorter amplicons, implying more efficient exon skipping than may actually be induced. Unless high fidelity RT-PCR systems are used, strand slippage during annealing/elongation steps will generate normal length transcripts that are artifacts of the amplification.
- Subjects :
- 0301 basic medicine
congenital, hereditary, and neonatal diseases and abnormalities
Messenger RNA
biology
Morpholino
Oligonucleotide
Computational biology
Amplicon
Exon skipping
In vitro
03 medical and health sciences
Exon
030104 developmental biology
0302 clinical medicine
biology.protein
Dystrophin
030217 neurology & neurosurgery
Subjects
Details
- ISBN :
- 978-1-4939-8650-7
- ISBNs :
- 9781493986507
- Database :
- OpenAIRE
- Journal :
- Methods in Molecular Biology ISBN: 9781493986507
- Accession number :
- edsair.doi...........b79515a30b77e73d04efd0ed3ada7855