优化了CRISPR/Cas9系统,用于的DF1细胞中的基因淘汰.
Kexin Zou1, Fang Wang1, Zechun Zhang1
1College of Animal Science and Technology, Northwest A&F University, Yangling, Shaanxi 712100, China.
Poultry science
|August 10, 2023
概括
在细胞中提高了CRISPR/Cas9基因编辑效率,使用了替代记者. 双 sgRNAs 增加了片段删除,而安普利康序列测定被证明比 T7E1 试验更为敏感,用于评估编辑结果.
科学领域:
- 分子生物学分子生物学
- 遗传学 是一个遗传学.
- 生物技术是生物技术.
背景情况:
- 在家禽中CRISPR技术的实用性受到编辑效率的限制.
- 了解影响细胞中CRISPR/Cas9基因淘汰效率的因素至关重要.
研究的目的:
- 为了研究影响DF1细胞中CRISPR/Cas9基因淘汰效率的因素.
- 为了比较基因组编辑评估方法和分析indel资料.
主要方法:
- 设计的单个或双个sgRNA针对特定的基因外子 (AMH,TGFBR2,PPARG).
- 在的DF1细胞中利用了sgRNA-CRISPR/Cas9和记者载体.
- 使用T7E1和安普利康序列测试评估编辑效率.
主要成果:
- 替代记者系统提高了编辑效率,增加了选择后的独立比例.
- 安普利康测序显示出比T7E1测试更高的灵敏度来检测裂纹.
- 双 sgRNA 与单个 sgRNA 相比,增加了片段删除比率.
结论:
- 替代记者有效地促进了细胞中的CRISPR/Cas9编辑效率.
- 片测序是一种比T7E1测试更敏感的方法来评估CRISPR/Cas9向效率.
- 双重sgRNA可以增强片段删除,为家禽的特定基因组修改提供了一种策略.
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