从单个细胞中转移的元素的长读RNA测序使用CELLO-seqq
Sophie A Marlow1, Lauryn A Deaville1,2, Rebecca V Berrens3
1Department of Biochemistry, Oxford University, Oxford, UK.
Nature protocols
|July 16, 2025
概括
可转移元素 (TE) 在基因组中非常丰富. 一种新的方法,CELLO-seq,使用长读RNA测序来准确地绘制单细胞中的TE表达,从而提高我们对它们作用的理解.
科学领域:
- 基因组学就是基因组学.
- 分子生物学分子生物学
- 文字转录学 (Transcriptomics) 是一个学科.
背景情况:
- 可转移元素 (TE) 占哺乳动物基因组的50%.
- TE表达与发育和疾病有关.
- 短读测序在绘制高度相似的TE序列方面遇到了困难.
研究的目的:
- 为单细胞长读RNA测序 (CELLO-seq) 提出一种新的协议.
- 为了能够准确地绘制可转移元素 (TE) 衍生的读取到独特的基因组位置.
- 为了促进基因和TE异型表达在单细胞中的高可靠性分析.
主要方法:
- 开发CELLO-seq,一个单细胞长读RNA测序协议.
- 整合长的独特分子标识符 (UMI) 来纠正错误.
- 高聚合酶连锁反应 (PCR) 重复数量以提高数据的准确性.
主要成果:
- CELLO-seq允许高保真地映射TE衍生的读数到独特的位置.
- 该协议可以准确量化年轻,非常相似的TE的表达.
- CELLO-seq可以区分基因和TE异型.
结论:
- CELLO-seq克服了用于TE分析的短读序列的局限性.
- 这种方法允许深入探索单细胞内TE基因相互作用.
- 该协议是可访问的,可以在一周内完成.
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