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在单细胞胚胎中通过Cas9/RNA介导基因向生成基因修饰的Cynomolgus子

Yuyu Niu¹, Bin Shen², Yiqiang Cui³

¹Yunnan Key Laboratory of Primate Biomedical Research, Kunming 650500, China; Kunming Biomed International and National Engineering Research Center of Biomedicine and Animal Science, Kunming 650500, China.

|February 4, 2014

概括

研究人员成功地使用CRISPR/Cas9基因编辑来创建转基因Cynomolgus子. 这种高效的方法精确地准基因,为非人类灵长类动物的疾病建模和治疗开发提供了新的可能性.

科学领域:

生物技术是生物技术.
遗传学是一个遗传学.
灵长类动物研究研究.

背景情况:

非人类灵长类动物是人类疾病研究的关键模型.
为研究产生转基因灵长类动物一直是个挑战.
克里斯普尔/卡斯9技术提供了精确的基因组编辑能力.

研究的目的:

在Cynomolgus子中应用CRISPR/Cas9技术进行精确的基因向.
建立一种有效的方法来产生转基因灵长类动物.
评估CRISPR/Cas9在灵长类动物生殖系编辑中的有效性和安全性.

主要方法:

将Cas9mRNA和单导向RNA (sgRNA) 联合注入单细胞阶段的Cynomolgus子胚胎中.
针对特定基因的向,包括Ppar-γ和Rag1.1.
对非目标突变进行全面分析.

主要成果:

在Cynomolgus子中成功和精确的基因向.
同时破坏两个目标基因 (Ppar-γ和Rag1) 在单个步骤.
没有观察到可检测的目标外突变.

结论:

与Cas9mRNA和sgRNA一起注入单细胞阶段胚胎是一种高效可靠的方法,用于产生基因修饰的Cynomolgus子.
这种方法显著提高了在生物医学研究中使用转基因灵长类动物的潜力.
克里斯普尔/Cas9有助于开发新的灵长类动物模型,用于研究人类疾病和测试疗法.

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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...

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.