George Dickson, Helen Foster, Takis Athanasopoulos, Capucine Trollet, Ian R. Graham, Keith Foster, Paul S. Sharp and Dominic J. Wells (2008) Codon and mRNA sequence optimization of microdystrophin transgenes improves expression and physiological outcome in dystrophic mdx mice following AAV2/8 gene transfer.. Molecular Therapy, 16 (11). pp. 1825–1832 . ISSN 1525-0016
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Duchenne muscular dystrophy is a fatal muscle wasting disorder. Lack of dystrophin compromises the integrity of the sarcolemma and results in myofibres that are highly prone to contraction induced injury. rAAV mediated dystrophin gene transfer strategies to muscle for the treatment of DMD have been limited by the small cloning capacity of rAAV vectors and high titres needed to achieve efficient systemic gene transfer. Here we assess the impact of codon optimisation on microdystrophin (ΔAB/R3-R18/ΔCT) expression and function in the mdx mouse and compare the function of two different configurations of codon-optimised microdystrophin genes (ΔAB/R3-R18/ΔCT & ΔR4- R23/ΔCT), under the control of a muscle restrictive promoter (Spc5-12). Codon optimisation of microdystrophin significantly increases levels of microdystrophin mRNA and protein following intramuscular and systemic administration of plasmid DNA or rAAV2/8. Physiological assessment demonstrates that codon optimisation of ΔAB/R3- R18/ΔCT results in significant improvement in specific force, but does not improve resistance to eccentric contractions compared to non-codon-optimised ΔAB/R3- R18/ΔCT. However, codon-optimised microdystrophin ΔR4-R23/ΔCT completely restored specific force generation and provided substantial protection from contraction induced injury. These results demonstrate that codon optimisation of microdystrophin under the control of a muscle specific promoter can significantly improve expression levels such that reduced titres of rAAV vectors will be required for efficient systemic administration.
This is a Draft version This version's date is: 02/09/2008 This item is peer reviewed
https://repository.royalholloway.ac.uk/items/2abe3570-3fde-7680-9749-1a268cd34717/1/
Deposited by () on 31-Mar-2010 in Royal Holloway Research Online.Last modified on 22-Dec-2010
(C) 2008 Elsevier, whose permission to mount this version for private study and research is acknowledged. The repository version is the author's final draft.
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