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通过长读纳米孔测序来探索细胞外囊的转录组多样性

Juan-Carlos A Padilla^1,2, Seda Barutcu¹, Gabrielle Deschamps-Francoeur¹

¹Institut de Recherches Cliniques de Montréal (IRCM), Montréal, QC, Canada.

Methods in molecular biology (Clifton, N.J.)

|February 3, 2025

概括

纳米孔长读测序使得在细胞外囊泡 (EVs) 内的全长RNA的分析成为可能,为细胞间通信提供了新的见解. 这种方法克服了用于全面EV RNA分析的短读序列的局限性.

科学领域:

生物化学生物化学
分子生物学分子生物学
基因组学就是基因组学.

背景情况:

细胞外囊泡 (EVs) 通过RNA载荷调解细胞间的通信.
短读测序片段RNA,限制EV转录组的分析.
在EV中完全长度RNA的存在和完整性尚未得到充分理解.

研究的目的:

通过纳米孔测序,介绍一项用于分析EV中全长RNA的协议.
克服传统RNA测序方法在EV研究中的局限性.
探索EV RNA货物的完整性和异型多样性.

主要方法:

细胞外囊泡 (EV) 的RNA净化.
补充DNA (cDNA) 库准备用于纳米孔测序.
牛津纳米孔技术 (ONT) MinION测序平台的使用.

主要成果:

纳米孔长读测序使得全长EVRNA的分析成为可能.
这种方法提供了对EV RNA完整性和异型多样性的更深入的见解.
使用ONT进行EV RNA测序的综合性协议详细说明.

结论:

长读纳米孔测序是EV RNA研究的强大工具.
这种方法增强了对EV介导细胞间通信的理解.
提出的协议有助于详细分析EV转录组.

关键词:

电动汽车 (EVs) 是一种电动汽车.细胞外囊泡中的细胞外囊泡.长读序列的测序方式纳米孔测序的测序牛津纳米孔技术公司在Poly-A中使用.多基化转录组的多基化转录组.有关RNA测序的RNA测序转录异型的异形.

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