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Updated: Jan 13, 2026

Retroviral Scanning: Mapping MLV Integration Sites to Define Cell-specific Regulatory Regions

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PRISM-Seq:一种超灵敏的测序方法,用于绘制隐形病毒集成部位的地图.

V K Pal, M Canis, E Stone

bioRxiv : the preprint server for biology

|January 9, 2026

概括

我们开发了PRISM-seq,这是一种超敏感的方法,用于绘制基因组中的逆转录病毒DNA插入的地图. 这种技术可以准确地识别整合部位,即使是从单个分子,提高基因疗法安全性和HIV-1研究.

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科学领域:

分子生物学分子生物学
基因组学就是基因组学.
病毒学病毒学

背景情况:

复原病毒融入宿主基因组对于HIV-1的持久性和在基因和细胞疗法中的lentiviral vector应用至关重要.
目前的整合场所测试在灵敏度,输入需求和分析复杂性方面存在局限性,缺乏单分子验证.
准确地绘制逆转录病毒融合部位的地图对于评估基因治疗的安全性和了解病毒动态至关重要.

研究的目的:

引入PRISM-seq,这是一个超敏感的工作流程,用于全基因组恢复lentiviral-host连接.
开发BulkIntSiteR,这是一个用于集成站点注释的自动化管道.
为了验证PRISM-seq在不同基因组区域和单分子分辨率上的性能.

主要方法:

PRISM-seq工作流程用于全基因组恢复lentiviral-host连接.
BulkIntSiteR开源管道用于自动集成网站注释.
测试和放大相关噪声的系统表征.
开发一个五步质量控制框架,以消除PCR和测序文物.

主要成果:

PRISM-seq准确地识别了各种基因组环境中的前病毒插入,包括欧克罗马丁,异性色素和中心区域.
高可信度集成事件被检测到一个单个输入模板分子.
与BulkIntSiteR相结合的PRISM-seq工作流,在大幅降低成本的情况下,表现与高输入试验相当或超过高输入试验.
一个强大的质量控制框架有效地消除了PCR和测序文物.

结论:

PRISM-seq提供了一种超灵敏且准确的方法,用于全基因组的复原病毒整合部位映射.
该工作流能够对集成事件进行高分辨率分析,这对于基因和细胞疗法开发和HIV-1研究至关重要.
PRISM-seq促进了定量克隆追踪,并为现有测试提供了具有成本效益的替代方案.