寡核酸中的合成错误通过同质导向修复传播到基因组中
Stacia K Wyman1, Zulema Romero2, Seok-Jin Heo3
1Innovative Genomics Institute, University of California, Berkeley, CA 94720, USA.
bioRxiv : the preprint server for biology
|July 17, 2025
概括
用于基因编辑的单链DNA寡核酸 (ssODNs) 的合成错误很常见,并因制造商而异. 这些错误在同质定向修复 (HDR) 过程中被纳入基因组,影响治疗策略.
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:
- 单链寡核酸 (ssODN) 是通过同质导向修复 (HDR) 来进行基因编辑的关键捐赠模板.
- 确保ssODNs的序列忠实性对于基因编辑应用的准确性和安全性至关重要.
研究的目的:
- 为了评估从不同制造商获得的ssODN的序列保真度.
- 为了确定ssODNs中的合成错误是否在HDR期间传播到基因组中.
主要方法:
- 来自三家商业制造商的ssODN的深度测序.
- 基因组DNA的深度测序在基因编辑后使用这些ssODN进行HDR.
主要成果:
- 在不同ssODN制造商中观察到单核酸和小删除合成错误的比例有显著差异.
- 发现ssODN模板中存在的这些合成错误通过HDR过程被纳入编辑的基因组.
结论:
- 商用ssODNs的序列保真性是可变的,并且包含合成错误.
- 同源性定向修复可以将这些ssODN合成错误传播到基因组中,这代表了基因编辑策略中的另一个错误来源.
- 治疗基因校正策略必须考虑这些ssODN衍生的错误,以确保安全性和有效性.
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