USING COMPLEMENTARY DNA FROM MyoD-TRANSDUCED FIBROBLASTS TO SEQUENCE LARGE MUSCLE GENES
Leigh B Waddell, Nicole Monnier, Sandra T Cooper, Kathryn N North, Nigel F Clarke
MUSCLE & NERVE | WILEY-BLACKWELL | Published : 2011
Large muscle genes are often sequenced using complementary DNA (cDNA) made from muscle messenger RNA (mRNA) to reduce the cost and workload associated with sequencing from genomic DNA. Two potential barriers are the availability of a frozen muscle biopsy, and difficulties in detecting nonsense mutations due to nonsense-mediated mRNA decay (NMD). We present patient examples showing that use of MyoD-transduced fibroblasts as a source of muscle-specific mRNA overcomes these potential difficulties in sequencing large muscle-related genes.
Awarded by NHMRC, Australia
The authors thank the study patients and their families for their invaluable contributions, and Dr. Mark Davis, Dr. Harriet Lo, Dr. Katrina Reardon, and Dr. Lisa Riley, who also contributed to this study. This work was supported by grants from the NHMRC, Australia (206529 and 571287 to N.F.C., 505004 to L.B.W., 301946 to S.T.C., and 301946 and 403941 to K.N.N.), and by the MDA of New South Wales (to N.F.C., K.N.N., S.T.C.), the MDA of the USA (to S.T.C.), the Brain Foundation Australia (to S.T.C.), and the Jain Foundation (to S.T.C.).