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用长时间读取的纳米孔测序对细胞外囊泡的多基化转录组进行剖析
Juan-Carlos A Padilla1,2, Seda Barutcu1, Ludovic Malet1
1Institut de Recherches Cliniques de Montréal (IRCM), 110 Avenue des Pins, Ouest, Montréal, QC, H2W 1R7, Canada.
BMC genomics
|September 22, 2023
概括
细胞外囊泡 (EVs) 含有多种全长RNA转录,包括mRNA和长非编码RNA. 纳米孔测序揭示了这些EV RNA配置文件,为它们的组成提供了新的见解.
科学领域:
- 分子生物学分子生物学
- 基因组学就是基因组学.
- 生物化学 生物化学
背景情况:
- 以前对细胞外囊泡 (EV) 转录组的研究经常使用RNA碎片化,限制了RNA完整性和异型多样性的分析.
- 假设EV相关的mRNAs主要是碎片化产物,对全长mRNA存在的理解有限.
研究的目的:
- 用长读纳米孔RNA测序来表征EVs的全长多基化 (多A) 转录组.
- 调查EV中全长RNA异型的存在和多样性.
主要方法:
- 从K562细胞中长时间读取EVs的纳米孔RNA测序.
- 分析RNA生物型,包括mRNA,长非编码RNA和伪基因.
- 在EV和整个细胞 (WC) 之间比较全长转录异形比率.
主要成果:
- 在EV中检测到443个丰富和280个枯竭的RNA.
- 确定了全长的转录,mRNA是EVs (58.13%) 和WC (43.93%) 中的主导生物型.
- 在EV中观察到各种各样的全长转录异型,反映但通常与细胞样本的比例不同.
结论:
- 这项研究为EV中多-A转录异型的组成多样性提供了新的见解.
- 突出了纳米孔测序的实用性,用于查询分泌囊泡的全长RNA含量.
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