Knockout of Myostatin by Zinc-finger Nuclease in Sheep Fibroblasts and Embryos
2
조회
0
다운로드
1
인용
0
참고문헌
키워드
초록
Myostatin (MSTN) can negatively regulate the growth and development of skeletal muscle, and natural mutations can cause “double-muscling” trait in animals. In order to block the inhibiting effect of MSTN on muscle growth, we transferred zinc-finger nucleases (ZFN) which targeted sheep MSTN gene into cultured fibroblasts. Gene targeted colonies were isolated from transfected fibroblasts by serial dilution culture and screened by sequencing. Two colonies were identified with mono-allele mutation and one colony with bi-allelic deletion. Further, we introduced the MSTN-ZFN mRNA into sheep embryos by microinjection. Thirteen of thirtyseven parthenogenetic embryos were targeted by ZFN, with the efficiency of 35%. Our work established the technical foundation for generation of MSTN gene editing sheep by somatic cloning and microinjection ZFN into embryos.
Abstract
Myostatin (MSTN) can negatively regulate the growth and development of skeletal muscle, and natural mutations can cause “double-muscling” trait in animals. In order to block the inhibiting effect of MSTN on muscle growth, we transferred zinc-finger nucleases (ZFN) which targeted sheep MSTN gene into cultured fibroblasts. Gene targeted colonies were isolated from transfected fibroblasts by serial dilution culture and screened by sequencing. Two colonies were identified with mono-allele mutation and one colony with bi-allelic deletion. Further, we introduced the MSTN-ZFN mRNA into sheep embryos by microinjection. Thirteen of thirtyseven parthenogenetic embryos were targeted by ZFN, with the efficiency of 35%. Our work established the technical foundation for generation of MSTN gene editing sheep by somatic cloning and microinjection ZFN into embryos.
참고문헌 (25)
- Bibikova, M, "Targeted chromosomal cleavage and mutagenesis in Drosophila using zinc-finger nucleases", Genetics, 161, pp.1169~1175 (2002)
- Boman, I. A., "A frameshift mutation in the coding region of the myostatin gene (MSTN) affects carcass conformation and fatness in Norwegian White Sheep (Ovis aries)", Anim. Genet, 40, pp.418~422 (2009)
- Chu, X., "Characterization of multidrug resistance 1a", P-glycoprotein knockout rats generated by zinc finger nucleases, 2012, pp.81, 220~227
- Clop, A., "A mutation creating a potential illegitimate microRNA target site in the myostatin gene affects muscularity in sheep", Nat. Genet, 38, pp.813~818 (2006)
- Cui, X, "Targeted integration in rat and mouse embryos with zinc-finger nucleases", Nat. Biotechnol, 29, pp.64~67 (2011)
- Doyon, Y., "Heritable targeted gene disruption in zebrafish using designed zincfinger nucleases", Nat. Biotechnol, 26, pp.702~708 (2008)
- Flisikowska, T., "Efficient immunoglobulin gene disruption and targeted replacement in rabbit using zinc finger nucleases", PLoS ONE, 6, pp.e21045~ (2011)
- Geurts, A. M, "Knockout rats via embryo microinjection of zinc-finger nucleases", Science, 325, pp.433~ (2009)
- Hauschild, J., "Efficient generation of a biallelic knockout in pigs using zinc-finger nucleases", Proc. Natl. Acad. Sci. USA, 108, pp.12013~12017 (2011)
- Hu, L. Y., "An alternative method for cDNA cloning from surrogate eukaryotic cells transfected with the corresponding genomic DNA", Biotechnol. Lett, 34, pp.1251~1255 (2012)
- Kambadur, R., "Mutations in myostatin (GDF8) in double-muscled Belgian Blue and Piedmontese cattle", Genome Res, 7, pp.910~916 (1997)
- Lai, L, "Production of alpha-1, 3-galactosyltransferase knockout pigs by nuclear transfer cloning", Science, 295, pp.1089~1092 (2002)
- Lloyd, A, "Targeted mutagenesis using zinc-finger nucleases in Arabidopsis", Proc. Natl. Acad. Sci. USA, 102, pp.2232~2237 (2005)
- McCreath, K. J., "Production of gene-targeted sheep by nuclear transfer from cultured somatic cells", Nature, 405, pp.1066~1069 (2000)
- McPherron, A. C., "Double muscling in cattle due to mutations in the myostatin gene", Proc. Natl. Acad. Sci. USA, 94, pp.12457~12461 (1997)
- Meng, X., "Targeted gene inactivation in zebrafish using engineered zinc-finger nucleases", Nat. Biotechnol, 26, pp.695~701 (2008)
- Meyer, M., "Gene targeting by homologous recombination in mouse zygotes mediated by zinc-finger nucleases", Proc. Natl. Acad. Sci. USA, 107, pp.15022~15026 (2010)
- Mosher, D. S, "A mutation in the myostatin gene increases muscle mass and enhances racing performance in heterozygote dogs", PLoS Genet, 3, pp.e79~ (2007)
- Park, S. J., "Oxamflatin improves developmental competence of porcine somatic cell nuclear transfer embryos", Cell. Reprogram, 14, pp.398~406 (2012)
- Patel, K., "The function of Myostatin and strategies of Myostatin blockade-new hope for therapies aimed at promoting growth of skeletal muscle", Neuromuscul. Disord, 15, pp.117~126 (2005)
- Richt, J. A., "Production of cattle lacking prion protein", Nat. Biotechnol, 25, pp.132~138 (2007)
- Takasu, Y., "Targeted mutagenesis in the silkworm Bombyx mori using zinc finger nuclease mRNA injection", Insect Biochem. Mol. Biol, 40, pp.759~765 (2010)
- Young, J. J., "Efficient targeted gene disruption in the soma and germ line of the frog Xenopus tropicalis using engineered zinc-finger nucleases", Proc. Natl. Acad. Sci. USA, 108, pp.7052~7057 (2011)
- Yu, S., "Highly efficient modification of beta-lactoglobulin (BLG) gene via zinc-finger nucleases in cattle", Cell Res, 21, pp.1638~1640 (2011)
- Zhang, C., "Targeted disruption of the sheep MSTN gene by engineered zinc-finger nucleases", Mol. Biol. Rep, 41, pp.209~215 (2014)