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Myostatin gene targeting in cultured China Han ovine myoblast cells

Published online by Cambridge University Press:  01 November 2007

L. Zhang ,
X. Yang ,
X. An  and
Y. Chen
L. Zhang
Affiliation:
State Key Laboratory for Agrobiotechnology, College of Biology, China Agricultural University, Yuanming Yuan West Road #2, Haidian District, 100094 Beijing, People’s Republic of China
X. Yang*
Affiliation:
State Key Laboratory for Agrobiotechnology, College of Biology, China Agricultural University, Yuanming Yuan West Road #2, Haidian District, 100094 Beijing, People’s Republic of China
X. An*
Affiliation:
State Key Laboratory for Agrobiotechnology, College of Biology, China Agricultural University, Yuanming Yuan West Road #2, Haidian District, 100094 Beijing, People’s Republic of China
Y. Chen
Affiliation:
State Key Laboratory for Agrobiotechnology, College of Biology, China Agricultural University, Yuanming Yuan West Road #2, Haidian District, 100094 Beijing, People’s Republic of China
*
E-mail: [email protected]
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Abstract

Myostatin (MSTN), a member of the transforming growth factor-β superfamily, has been shown to be a negative regulator of myogenesis. Natural mutation in beef cattle causes double-muscling phenotypes. We report an investigation designed to knockout the MSTN gene by gene targeting in ovine myoblast cells. Two promoter-trap targeting vectors MSTN-green fluorescent protein (GFP) and MSTN-neo were constructed and used to transfect foetal and neonatal ovine primary myoblast cells. Both GFP-expressing cells and drug-resistant cells were obtained. Targeted cells expressing GFP were confirmed by polymerase chain reaction (PCR) assay and drug-resistant cells were characterised by PCR and Southern blot after growing into cell clones.

Keywords

gene targetingmyoblast cellmyostatin transgenics
Type
Full Paper
Information
animal , Volume 1 , Issue 10 , November 2007 , pp. 1401 - 1408
Copyright
Copyright © The Animal Consortium 2007

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References

Denning, C, Burl, S, Ainslie, A, Bracken, J, Dinnyes, A, Fletcher, J, King, T, Ritchie, M, Ritchie, WA, Rollo, M, de Sousa, P, Travers, A, Wilmut, I, Clark, AJ 2001. Deletion of the α-1,3 galactosyl transferase (GGTA1) gene and the prion protein (PrP) gene in sheep. Nature Biotechnology 19, 559–562.CrossRefGoogle ScholarPubMed
Hanson, KD, Sedivy, JM 1995. Analysis of biological selection for high-efficiency gene targeting. Molecular and Cell Biology 15, 45–51.CrossRefGoogle ScholarPubMed
Joyner, AL, Skarnes, WC, Rossant, J 1989. Production of a mutation in mouse En-2 gene by homologous recombination in embryonic stem cells. Nature 338, 153–156.CrossRefGoogle ScholarPubMed
Kambadur, R, Sharma, M, Smith, TP, Bass, JJ 1997. Mutations in myostatin (GDF8) in double-muscled Belgian Blue and Piedmontese cattle. Genome Research 7, 910–915.CrossRefGoogle ScholarPubMed
Kaufman, SJ, Foster, RF 1988. Replicating myoblasts express a muscle-specific phenotype. Proceedings of the National Academy of Sciences 85, 9606–9610.CrossRefGoogle ScholarPubMed
McCreath, KJ, Howcroft, J, Campbell, KH, Colman, A, Schnieke, AE, Kind, AJ 2000. Production of gene-targeted sheep by nuclear transfer from cultured somatic cells. Nature 405, 1066–1069.CrossRefGoogle ScholarPubMed
McPherron, AC, Lawler, AM, Lee, SJ 1997. Regulation of skeletal muscle mass in mice by a new TGF-b superfamily member. Nature 387, 83–90.CrossRefGoogle ScholarPubMed
Rando, TA, Blau, HM 1994. Primary mouse myoblast purification, characterization, and transplantation for cell-mediated gene therapy. The Journal of Cell Biology 125, 1275–1287.CrossRefGoogle ScholarPubMed
Richler, C, Yaffe, D 1970. The in vitro cultivation and differentiation capacities of myogenic cell lines. Developmental Biology 23, 1–22.CrossRefGoogle ScholarPubMed
Schwartzberg, PL, Robertson, EJ, Goff, SP 1990. Targeted gene disruption of the endogenous c-abl locus by homologous recombination with DNA encoding a selectable fusion protein. Proceedings of the National Academy of Sciences 87, 3210–3214.CrossRefGoogle ScholarPubMed
Yang, J, Ratovitski, T, Brady, JP, Solomon, MB, Wells, KD, Wall, RJ 2001. Expression of myostatin pro domain results in muscular transgenic mice. Molecular Reproduction and Development 60, 351–361.CrossRefGoogle ScholarPubMed