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使用I-E型CRISPR-Cas3的基因组编辑在小鼠和老鼠的密细胞中

Kazuto Yoshimi¹, Akihiro Kuno², Yuko Yamauchi³

¹Division of Animal Genetics, Laboratory Animal Research Center, Institute of Medical Science, The University of Tokyo, Tokyo 108-8639, Japan; Division of Genome Engineering, Center for Experimental Medicine and Systems Biology, Institute of Medical Science, University of Tokyo, Tokyo 108-8639, Japan.

Cell reports methods

|August 9, 2024

概括

I型CRISPR-Cas3系统能够在小鼠和老鼠的阴茎中进行高效的大规模基因组编辑,在没有目标突变的情况下产生大量的删除. 这一进步扩大了基因工程和模型动物生成的可能性.

科学领域:

分子生物学分子生物学
遗传学遗传学是一个
生物技术是生物技术.

背景情况:

I型CRISPR系统是基因组修改的强大工具.
它在胚胎编辑中用于创建模型动物的应用以前是未确定的.

研究的目的:

通过I-E型CRISPR-Cas3系统,在阴囊中演示基因组编辑.
在模型动物中建立基因工程的多功能方法.

主要方法:

利用I-E型CRISPR-Cas3系统在小鼠和老鼠中进行胚胎编辑.
采用长期读取的基于测序的多重基因型鉴定方法来检测删除.
在基因编辑中应用了囊电穿孔方法.

主要成果:

在小鼠的目标位点 (效率为40-70%) 实现了有效的缺失生成 (几千个基对).
在小鼠和大鼠中证明了成功的基因组编辑,包括囊胞电穿孔.
在编辑的小鼠中,没有确认的非目标突变.
在小鼠中使用捐赠体等离子体成功进行了SNP交换和基因组替代.

结论:

I型CRISPR-Cas3系统对于在紫体中进行大规模的基因组编辑是有效的.
这种方法提供了更大的灵活性和更广泛的应用在基因工程跨物种.
开发的技术有助于生成转基因模型动物.

关键词:

CP: 生物技术生物技术科普:遗传学在CP:遗传学.这就是CRISPR-Cas3.电穿孔的电穿孔是什么意思基因组编辑基因组编辑在敲门的时候.淘汰赛淘汰赛是一个淘汰赛.大量的删除删除.这里是鼠标鼠标鼠标鼠标鼠标鼠标.多重基因型定型多重基因型定型鼠标鼠标鼠标鼠标鼠标替代品替代品替代品替代品胚胎细胞 (zygote) 是一个细胞.

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