基因工程小鼠的基因定型协议
Advait Limaye1, Kyoungin Cho2, Bradford Hall1
1National Institute of Dental and Craniofacial Research, National Institutes of Health, Bethesda, Maryland.
Current protocols
|November 20, 2023
概括
本指南详细介绍了识别转基因小鼠模型的方法,重点是聚合酶连锁反应 (PCR) 技术,用于传统和集群定期间隔短平行列重复 (CRISPR) 基因编辑. 它为研究环境中准确的遗传特征验证提供了必要的协议.
科学领域:
- * 遗传学和基因组学
- * 哺乳动物模型系统
- * 分子生物学 * 分子生物学
背景情况:
- *实验室小鼠对于研究基因功能和人类疾病建模至关重要.
- *像CRISPR这样的基因编辑技术的进步使得精确的小鼠基因组操纵成为可能.
- *标准化方法对于在研究设施中一致识别转基因小鼠模型至关重要.
研究的目的:
- * 为识别转基因小鼠模型提供全面的指导方针.
- *详细介绍终点聚合酶链反应 (PCR) 基因定型方法.
- * 概述用于识别使用新基因编辑技术创建的模型的策略,包括CRISPR.
主要方法:
- *从各种小鼠组织和样本中提取基因组DNA (尾巴活检,耳孔,胚囊,精液,血液,口腔抹布).
- * 终点聚合酶链反应 (PCR) 用于例行基因型定型.
- *包括T7E1/Surveyor在内的CRISPR特定测定,非目标突变检测和基因敲入/删除分析.
主要成果:
- * 建立了可靠从各种生物样本中净化DNA的协议.
- * 证明了终点PCR对转基因小鼠常规基因定型的有效性.
- * 展示了验证CRISPR中介编辑的方法,包括插入/删除检测和目标外分析.
结论:
- * 标准化分子技术对于精确描述转基因小鼠模型至关重要.
- *终点PCR仍然是基因型定型的强大方法,并补充了CRISPR编辑模型的专门测试.
- *这些指导方针促进使用转基因小鼠进行一致和可重复的研究.
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