GONAD: A Novel CRISPR/Cas9 Genome Editing Method that Does Not Require Ex Vivo Handling of Embryos.
GONAD: A Novel CRISPR/Cas9 Genome Editing Method that Does Not Require Ex Vivo Handling of Embryos.
复制标题
性腺：一种新型的CRISPR/CAS9基因组编辑方法，不需要离体处理胚胎。
DOI：
10.1002/0471142905.hg1508s88
复制
发表时间：
2016-01-01
影响因子：
--
通讯作者：
Ohtsuka M
中科院分区：
文献类型：
--
作者：
Gurumurthy CB;Takahashi G;Wada K;Miura H;Sato M;Ohtsuka M
用于在动物中产生所需基因组变化的转基因技术涉及三个关键步骤：分离受精卵，将转基因DNA显微注射到它们中，然后将其转移到受体雌性中。这些离体步骤已经广泛使用了30多年，它们也很容易适应最新的基因组编辑技术，如ZFN，TALEN和CRISPR/Cas9系统。我们最近开发了一种称为GONAD（通过输卵管核酸递送进行基因组编辑）的方法，该方法不需要转基因的所有三个关键步骤，因此缓解了广泛使用的动物转基因技术的瓶颈。这里我们提供GONAD系统的协议。
Transgenic technologies used for creating a desired genomic change in animals involve three critical steps: isolation of fertilized eggs, microinjection of transgenic DNA into them and their subsequent transfer to recipient females. These ex vivo steps have been widely used for over 3 decades and they were also readily adapted for the latest genome editing technologies such as ZFNs, TALENs and CRISPR/Cas9 systems. We recently developed a method called GONAD (Genome editing via Oviductal Nucleic Acids Delivery) that does not require all the three critical steps of transgenesis and therefore relieves the bottlenecks of widely used animal transgenic technologies. Here we provide protocols for the GONAD system.