用CRISPR/Cas9系统进行瘤性染色体重组的体内工程
Danilo Maddalo1, Eusebio Manchado1, Carla P Concepcion2
1Memorial Sloan Kettering Cancer Center, Cancer Biology and Genetics Program, 1275 York Avenue, New York, New York 10065, USA.
Nature
|October 23, 2014
概括
研究人员开发了一种新的CRISPR/Cas9方法,通过诱导特定的染色体重组来创建癌症的小鼠模型. 这种技术成功地生成了像4 (EML4) - 形淋巴瘤激酶 (ALK) 肺癌这样的状体微管相关蛋白的模型,该模型对ALK抑制剂做出了反应.
科学领域:
- 在瘤学瘤学.
- 遗传学 是一个遗传学.
- 分子生物学分子生物学
背景情况:
- 染色体重组是人类癌症发病的关键驱动因素,经常导致治疗性向的基因融合.
- 像4 (EML4) - 形淋巴瘤激酶 (ALK) 融合基因这样的体微管相关蛋白质,由染色体2的反转产生的,是非小细胞肺癌 (NSCLC) 的子集中的重要瘤基因.
- 在小鼠中建模这种遗传事件的现有方法是复杂的,需要广泛的生殖线操纵.
研究的目的:
- 使用CRISPR/Cas9技术建立一种高效的体内方法来诱导特定的染色体重排.
- 为了生成一种针对EML4-ALK驱动的肺癌的新型小鼠模型.
- 验证该模型在研究癌症病原和治疗反应方面的实用性.
主要方法:
- 利用CRISPR/Cas9系统的病毒介导输送来诱导成年小鼠体细胞中的向染色体逆转.
- 产生了一种专门含有EML4-ALK融合基因的小鼠模型.
- 分析了由此产生的瘤的组织病理学和分子特征,并评估了它们对ALK抑制剂的反应.
主要成果:
- 在体内,CRISPR/Cas9方法有效地诱导了特定的EML4-ALK逆转.
- 生成的小鼠模型准确地回顾了人类ALK阳性NSCLC的关键特征,包括瘤形成和融合基因的表达.
- 鼠标模型中的瘤对ALK抑制剂表现出敏感性,反映了人类患者的临床观察.
结论:
- 病毒介导的CRISPR/Cas9传递为模拟染色体重组和相关癌症 in vivo提供了一种有效的策略.
- 这种方法显著提高了为人类癌症创造基因工程小鼠模型的能力.
- 开发的EML4-ALK肺癌模型是针对ALK驱动的NSCLC进行临床前研究和药物开发的宝贵平台.
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