Genetic characterization and improved genotyping of the dysferlin-deficient mouse strain Dysf tm1Kcam

Skeletal Muscle

Michael Sinnreich²  Kevin P. Campbell¹  Kazuhiro Kobuke¹  Jochen Kinter²  Tatiana Wiktorowicz² 

[1] Howard Hughes Medical Institute, Departments of Molecular Physiology and Biophysics, Neurology, and Internal Medicine, The University of Iowa Roy J. and Lucille A. Carver College of Medicine, Iowa City 52242, IA, USA;Neuromuscular Research Group, Departments of Neurology and Biomedicine, University and University Hospital Basel, Petersgraben 4, Basel, 4031, Switzerland

关键词: Knock-out;    Targeted deletion;    Dysferlinopathies;    Genotyping protocol;    Dysferlin mouse models;   

Others  :  1231175
DOI  :  10.1186/s13395-015-0057-3

 received in 2015-05-04, accepted in 2015-09-03,  发布年份 2015

【 摘 要 】

Background

Mouse models of dysferlinopathies are valuable tools with which to investigate the pathomechanisms underlying these diseases and to test novel therapeutic strategies. One such mouse model is the Dysf ^tm1Kcam strain, which was generated using a targeting vector to replace a 12-kb region of the dysferlin gene and which features a progressive muscular dystrophy. A prerequisite for successful animal studies using genetic mouse models is an accurate genotyping protocol. Unfortunately, the lack of robustness of currently available genotyping protocols for the Dysf ^tm1Kcam mouse has prevented efficient colony management. Initial attempts to improve the genotyping protocol based on the published genomic structure failed. These difficulties led us to analyze the targeted locus of the dysferlin gene of the Dysf ^tm1Kcam mouse in greater detail.

Methods

In this study we resequenced and analyzed the targeted locus of the Dysf ^tm1Kcam mouse and developed a novel PCR protocol for genotyping.

Results

We found that instead of a deletion, the dysferlin locus in the Dysf ^tm1Kcam mouse carries a targeted insertion. This genetic characterization enabled us to establish a reliable method for genotyping of the Dysf ^tm1Kcam mouse, and thus has made efficient colony management possible.

Conclusion

Our work will make the Dysf ^tm1Kcam mouse model more attractive for animal studies of dysferlinopathies.

【 授权许可】

   
2015 Wiktorowicz et al.

【 图 表 】

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