基于板块的10X基因组学兼容的基于智能seq3xpress的单细胞RNA测序.
Koen Deserranno1, Elise Callens1, Danique Berrevoet1
1Laboratory of Pharmaceutical Biotechnology - NXTGNT, Faculty of Pharmaceutical Sciences, Ghent University, Ottergemsesteenweg 460, BLOK A, FLOOR3, Ghent, 9000, Belgium.
BMC genomics
|December 6, 2025
概括
一种新的基于板式的方法 (PB10X) 提高了单细胞RNA测序 (scRNA-seq) 与10X基因组学技术的兼容性. 这种具有成本效益的方法改善了免疫受体谱系测序和基因表达概况,特别是在小细胞群中.
科学领域:
- 单细胞转录组学 单细胞转录组学
- 基因组学就是基因组学.
- 免疫学 免疫学 免疫学
背景情况:
- 10X Genomics (10X) 是一个领先的单细胞RNA测序 (scRNA-seq) 技术,用于基因表达和T细胞受体 (TCR) 分析.
- 它基于微流体的方法限制了索引分类与scRNA-seq数据的直接配对,并且对于小细胞群体来说是昂贵的.
研究的目的:
- 开发一个基于板块的,10X兼容 (PB10X) scRNA-seq策略.
- 克服现有的基于微流体的scRNA-seq方法的局限性,以成本有效地对小细胞样本进行分析.
主要方法:
- 根据智能seq3xpress (SS3X) 原则开发了一个基于板块的,兼容10X (PB10X) 的scRNA-seq策略.
- 利用光激活细胞分类 (FACS) 进行直接对384个井板进行索引分类.
- 生成的cDNA兼容10X单细胞5'库套件 (V(D) J和基因表达).
主要成果:
- 在Jurkat淋巴细胞上,PB10X在V(D) J和基因表达分析中表现出高性能.
- 在有限的测序深度下,为81.71%的细胞实现了对TCRαβ链的配对测序.
- 检测到平均每细胞4343个基因和16137个UMI,具有高的独特映射读数和蛋白质编码基因.
- 解决了在SS3X.中观察到的丝入侵文物.
结论:
- 新的PB10X方法提供了具有成本效益的,基于板式的10X兼容性免疫受体谱和基因表达造型.
- PB10X是一种有前途的单细胞转录组学策略,特别适用于细胞数量有限的免疫学研究.
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