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在人类胚胎中Cas9裂变后的异构特异性染色体去除

Michael V Zuccaro¹, Jia Xu², Carl Mitchell³

¹Department of Pediatrics and Naomi Berrie Diabetes Center, Columbia University, New York, NY 10032, USA; Columbia University Stem Cell Initiative, New York, NY 10032, USA.

|October 30, 2020

概括

人类胚胎中的基因编辑可以纠正导致遗传性疾病的突变. 然而,这项研究显示,常见的修复机制会导致意外的染色体损失和缺陷,这对安全的临床应用构成重大挑战.

科学领域:

人类胚胎学
分子遗传学
基因编辑技术

背景情况:

遗传性疾病给家庭带来很大的负担.
目前的生育治疗通常依赖于胚胎选择以避免遗传疾病.
基因编辑为纠正人类胚胎中致病突变提供了潜在的替代方案.

研究的目的:

在人类胚胎中评估Cas9诱导的双链断裂 (DSB) 的修复结果.
评估基因编辑对导致失明的特定突变的有效性和安全性.
确定基因编辑在人类早期发展的潜在挑战和意外后果.

主要方法:

在人类胚胎中引发了双链断裂 (DSB) 在父亲染色体上的EYS位点.
分析DNA修复结果,包括微同质介导末端连接 (MMEJ) 和非同质末端连接 (NHEJ).
评估Cas9异位裂变及其对染色体完整性的影响.

主要成果:

主要的修复结果是微同质介导的末端连接 (MMEJ),导致第一个细胞周期中的读取框架的非马赛克恢复.
大约一半的诱导断裂没有得到修复,导致无法检测到的父性基因和随后的染色体臂损失.
由于双性裂变,Cas9的脱导致染色体损失和半性插入/删除 (indels).

结论:

虽然基因编辑可以恢复人类胚胎的读取框架,但像MMEJ和未修复的断裂这样的意外修复结果很常见.
Cas9诱导的DSB可以导致显著的染色体不稳定性,包括损失和缺陷,对临床应用构成风险.
在人类胚胎中实现安全有效的基因突变纠正仍然存在重大挑战,需要进一步研究精确的修复机制和非目标效应.

关键词:

其他国家染色体损失双链断裂的修复胚胎干细胞生殖基因编辑同类重组人类胚胎在同类基因间进行重组微同质介导的末端连接细胞分裂

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