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A PCR‐based assay for the wild‐type dystrophin gene transferred into mdx mouse
Author(s) -
Shrager Joseph B.,
Naji Ali,
Kelly Alan M.,
Stedman Hansell H.
Publication year - 1992
Publication title -
muscle and nerve
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.025
H-Index - 145
eISSN - 1097-4598
pISSN - 0148-639X
DOI - 10.1002/mus.880151012
Subject(s) - dystrophin , mdx mouse , duchenne muscular dystrophy , primer (cosmetics) , muscular dystrophy , biology , microbiology and biotechnology , myocyte , gene , polymerase chain reaction , real time polymerase chain reaction , genetics , chemistry , organic chemistry
Myoblast transfer has emerged as a promising treatment for inherited myopathies such as Duchenne muscular dystrophy (DMD). Further development of the technique's therapeutic potential requires an experimental system in which issues of graft rejection can be clearly discriminated from those related to myoblast biology. Here we report the development and initial application of a quantitative assay for myogenic cells bearing a wild‐type dystrophin gene following transfer into the mdx mouse. The technique relies upon the ability of a mutagenizing polymerase chain reaction (PCR) primer to create a new restriction site in the amplification production of the wild‐type, but not the mdx dystrophin gene. The ratio of host to donor cells can be determined from muscle biopsies as small as 1 mg, regardless of donor H‐2 background. This simple technique should allow a number of basic questions related to myoblast and direct gene transfer to be addressed using the mdx mouse model. © 1992 John Wiley & Sons, Inc.