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Somatic cell reprogramming-free generation of genetically modified pigs

Genetically modified pigs for biomedical applications have been mainly generated using the somatic cell nuclear transfer technique; however, this approach requires complex micromanipulation techniques and sometimes increases the risks of both prenatal and postnatal death by faulty epigenetic reprogr...

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Detalles Bibliográficos
Autores principales: Tanihara, Fuminori, Takemoto, Tatsuya, Kitagawa, Eri, Rao, Shengbin, Do, Lanh Thi Kim, Onishi, Akira, Yamashita, Yukiko, Kosugi, Chisato, Suzuki, Hitomi, Sembon, Shoichiro, Suzuki, Shunichi, Nakai, Michiko, Hashimoto, Masakazu, Yasue, Akihiro, Matsuhisa, Munehide, Noji, Sumihare, Fujimura, Tatsuya, Fuchimoto, Dai-ichiro, Otoi, Takeshige
Formato: Online Artículo Texto
Lenguaje:English
Publicado: American Association for the Advancement of Science 2016
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5023319/
https://www.ncbi.nlm.nih.gov/pubmed/27652340
http://dx.doi.org/10.1126/sciadv.1600803
Descripción
Sumario:Genetically modified pigs for biomedical applications have been mainly generated using the somatic cell nuclear transfer technique; however, this approach requires complex micromanipulation techniques and sometimes increases the risks of both prenatal and postnatal death by faulty epigenetic reprogramming of a donor somatic cell nucleus. As a result, the production of genetically modified pigs has not been widely applied. We provide a simple method for CRISPR (clustered regularly interspaced short palindromic repeats)/Cas9 gene editing in pigs that involves the introduction of Cas9 protein and single-guide RNA into in vitro fertilized zygotes by electroporation. The use of gene editing by electroporation of Cas9 protein (GEEP) resulted in highly efficient targeted gene disruption and was validated by the efficient production of Myostatin mutant pigs. Because GEEP does not require the complex methods associated with micromanipulation for somatic reprogramming, it has the potential for facilitating the genetic modification of pigs.