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Rui Tian¹, Chen Cao², Dan He³

¹Generulor Co., Ltd., Zhuhai 519000, Guangdong, China.

Med (New York, N.Y.)

|June 6, 2023

概括

一种新的方法,通过测序 (AID-seq) 进行适配器介导的目标外识别,在CRISPR基因组编辑中准确检测低频的目标外突变. 这种敏感的技术有助于开发更安全的基因疗法,通过识别最佳指导RNA和表征新型CRISPR系统.

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:

克里斯普尔基因组编辑具有显著的临床潜力,但面临着非目标效应的挑战.
准确检测这些非目标突变对于安全的临床转换至关重要.

研究的目的:

开发一种敏感和特定的方法来检测CRISPR基因组编辑中的低频脱突变.
建立一个高通量平台,用于选择最佳导向RNA和表征新型CRISPR核酶.

主要方法:

通过测序 (AID-seq) 开发了适配器介导的目标外识别,用于全面的目标外检测.
与AID-seq一起实施一个聚合策略,同时分析多个导向RNA.
利用深度学习 (CRISPR-Net) 来构建一个目标外预测模型.

主要成果:

AID-seq在检测来自各种CRISPR核酶 (Cas9,Cas12a) 的脱突变方面表现出高灵敏度和特异性.
合并策略成功选了抗病毒治疗的高效和安全的标,并对一种新的CRISPR (FrCas9) 进行了分析.
在预测非目标效应方面,CRISPR-Net模型取得了高性能 (AUROC = 0.97,AUPRC = 0.29).

结论:

AID-seq代表了目前可用的最敏感和最具特异性的体外目标外检测方法.
聚合的AID-seq策略为优化CRISPR应用程序和发现新系统提供了一个快速,高吞吐量平台.

关键词:

在 AID-seqq.克里斯普尔脱离目标基础研究基础研究高通量目标外检测检测器在体外的目标外检测.目标之外的预测模型聚合了 AID-seqq 的数据.在 sgRNA 查中使用 sgRNA 查.

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