Somatic cell reprogramming-free generation of genetically modified pigs.

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 Adv

journal_title

Science advances

authors

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 T

doi

10.1126/sciadv.1600803

subject

Has Abstract

pub_date

2016-09-14 00:00:00

pages

e1600803

issue

9

issn

2375-2548

pii

1600803

journal_volume

2

pub_type

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