Abstract:
: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.
journal_name
Sci Advjournal_title
Science advancesauthors
Tanihara F,Takemoto T,Kitagawa E,Rao S,Do LT,Onishi A,Yamashita Y,Kosugi C,Suzuki H,Sembon S,Suzuki S,Nakai M,Hashimoto M,Yasue A,Matsuhisa M,Noji S,Fujimura T,Fuchimoto D,Otoi Tdoi
10.1126/sciadv.1600803subject
Has Abstractpub_date
2016-09-14 00:00:00pages
e1600803issue
9issn
2375-2548pii
1600803journal_volume
2pub_type
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