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Knockout of Myostatin by Zinc-finger Nuclease in Sheep Fibroblasts and Embryos
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...
Autores principales: | , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Asian-Australasian Association of Animal Production Societies (AAAP) and Korean Society of Animal Science and Technology (KSAST)
2016
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5003977/ https://www.ncbi.nlm.nih.gov/pubmed/27189642 http://dx.doi.org/10.5713/ajas.16.0130 |
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author | Zhang, Xuemei Wang, Liqin Wu, Yangsheng Li, Wenrong An, Jing Zhang, Fuchun Liu, Mingjun |
author_facet | Zhang, Xuemei Wang, Liqin Wu, Yangsheng Li, Wenrong An, Jing Zhang, Fuchun Liu, Mingjun |
author_sort | Zhang, Xuemei |
collection | PubMed |
description | 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 thirty-seven 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. |
format | Online Article Text |
id | pubmed-5003977 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2016 |
publisher | Asian-Australasian Association of Animal Production Societies (AAAP) and Korean Society of Animal Science and Technology (KSAST) |
record_format | MEDLINE/PubMed |
spelling | pubmed-50039772016-10-01 Knockout of Myostatin by Zinc-finger Nuclease in Sheep Fibroblasts and Embryos Zhang, Xuemei Wang, Liqin Wu, Yangsheng Li, Wenrong An, Jing Zhang, Fuchun Liu, Mingjun Asian-Australas J Anim Sci Article 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 thirty-seven 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. Asian-Australasian Association of Animal Production Societies (AAAP) and Korean Society of Animal Science and Technology (KSAST) 2016-10 2016-05-12 /pmc/articles/PMC5003977/ /pubmed/27189642 http://dx.doi.org/10.5713/ajas.16.0130 Text en Copyright © 2016 by Asian-Australasian Journal of Animal Sciences This is an open-access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/3.0/), which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited. |
spellingShingle | Article Zhang, Xuemei Wang, Liqin Wu, Yangsheng Li, Wenrong An, Jing Zhang, Fuchun Liu, Mingjun Knockout of Myostatin by Zinc-finger Nuclease in Sheep Fibroblasts and Embryos |
title | Knockout of Myostatin by Zinc-finger Nuclease in Sheep Fibroblasts and Embryos |
title_full | Knockout of Myostatin by Zinc-finger Nuclease in Sheep Fibroblasts and Embryos |
title_fullStr | Knockout of Myostatin by Zinc-finger Nuclease in Sheep Fibroblasts and Embryos |
title_full_unstemmed | Knockout of Myostatin by Zinc-finger Nuclease in Sheep Fibroblasts and Embryos |
title_short | Knockout of Myostatin by Zinc-finger Nuclease in Sheep Fibroblasts and Embryos |
title_sort | knockout of myostatin by zinc-finger nuclease in sheep fibroblasts and embryos |
topic | Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5003977/ https://www.ncbi.nlm.nih.gov/pubmed/27189642 http://dx.doi.org/10.5713/ajas.16.0130 |
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