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Updated: Jun 26, 2025

A Protocol for Multiple Gene Knockout in Mouse Small Intestinal Organoids Using a CRISPR-concatemer

A Protocol for Multiple Gene Knockout in Mouse Small Intestinal Organoids Using a CRISPR-concatemer

Published on: July 12, 2017

使用多目标CRISPR-Cas9系统生成一个器官缺陷动物模型

Jonathan Jun-Yong Lim^1,2, Yamato Murata¹, Shunsuke Yuri¹

¹Laboratory of Organ Developmental Engineering, Division of Biological Science, Graduate School of Science and Technology, Nara Institute of Science and Technology, 8916-5 Takayama-cho, Ikoma, Nara, 630-0912, Japan.

Scientific reports

|May 9, 2024

概括

这项研究引入了一种新的CRISPR-Cas9系统,用于创建器官缺陷的小鼠胚胎,从而使得跨物种金像体的产生成为可能. 这一突破有助于在小鼠模型中开发小鼠胸腺,用于研究.

科学领域:

发展生物学发展生物学
遗传学遗传学是一个
生物技术是生物技术.

背景情况:

胚胎细胞补充的基因淘汰动物模型往往缺乏可行性.
同性卵性突变胚胎很少,限制了研究潜力.

研究的目的:

开发一种使用CRISPR-Cas9.9生成器官缺陷胚胎的可靠方法.
为器官发育研究创建可行的跨物种金像体.

主要方法:

利用CRISPR-Cas9-sgRNAms系统在小鼠胚胎中诱导向细胞死亡.
在Foxn1的推动下,开发了一个使用Cas9的小鼠模型 (Foxn1Cas9; Rosa26_ms).
采用了与大鼠胚胎干细胞的胚胎细胞补充.

主要成果:

在实验室中,CRISPR-Cas9-sgRNAms系统有效诱导了细胞剥离.
产生的小鼠胚胎与一个无血型的表型.
成功创建了一个与老鼠细胞衍生的胸腺的跨物种金像体.

结论:

克里斯普尔-Cas9-sgRNAms系统为产生器官缺陷胚胎提供了一个可行的策略.
这种方法克服了传统的胚胎细胞补充基因淘汰模型的局限性.
能够使奇梅拉中不同物种的功能性器官发育.

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Efficient Production and Identification of CRISPR/Cas9-generated Gene Knockouts in the Model System Danio rerio

Efficient Production and Identification of CRISPR/Cas9-generated Gene Knockouts in the Model System Danio rerio

Published on: August 28, 2018

Removal of an Internal Translational Start Site from mRNA While Retaining Expression of the Full-Length Protein

Removal of an Internal Translational Start Site from mRNA While Retaining Expression of the Full-Length Protein

Published on: March 16, 2022

相关概念视频

In-vitro Mutagenesis

In-vitro Mutagenesis

To learn more about the function of a gene, researchers can observe what happens when the gene is inactivated or “knocked out,” by creating genetically engineered knockout animals. Knockout mice have been particularly useful as models for human diseases such as cancer, Parkinson’s disease, and diabetes.

CRISPR

CRISPR

Genome editing technologies allow scientists to modify an organism’s DNA via the addition, removal, or rearrangement of genetic material at specific genomic locations. These types of techniques could potentially be used to cure genetic disorders such as hemophilia and sickle cell anemia. One popular and widely used DNA-editing research tool that could lead to safe and effective cures for genetic disorders is the CRISPR-Cas9 system. CRISPR-Cas9 stands for Clustered Regularly Interspaced...