在人类细胞中进行基因表达,细胞表面蛋白质,突变状态和RNA拼接的多omics概况的GoT-Splice协议
Saravanan Ganesan1, Mariela Cortés-López1, Ariel D Swett1
1New York Genome Center, New York, NY 10013, USA; Division of Hematology and Medical Oncology, Department of Medicine and Meyer Cancer Center, Weill Cornell Medicine, New York, NY 10065, USA.
STAR protocols
|March 21, 2024
概括
转录组 (GoT) -Splice的基因定型使单细胞中精确的RNA拼接分析成为可能. 这种方法克服了识别突变和从测序数据中重建拼接信号的挑战.
科学领域:
- 分子生物学分子生物学
- 基因组学就是基因组学.
- 一个单细胞分析.
背景情况:
- 由于区分细胞类型和从短读序列中重建拼接信号的挑战,研究RNA拼接因子突变是困难的.
- 现有的方法缺乏准确分析复杂生物系统的剪接变化所需的精度.
研究的目的:
- 介绍转录组 (GoT) -Splice的基因型定型,这是一种用于精确RNA拼接分析的新协议.
- 克服区分野生类型和突变细胞的局限性,改善拼接信号重建.
主要方法:
- 结合了转录组 (GoT) 的基因型定型与长时间读取的单细胞转录组和细胞表面蛋白质组概况.
- 详细说明了长读库准备,cDNA重新放大,突变丰富,样本索引和GoT库准备的步骤.
主要成果:
- GoT-Splice协议有效地区分复杂组织中的野生类型和突变细胞.
- 它提供了拼接信号的准确重建,克服了短读序列的限制.
结论:
- 在单细胞水平上研究RNA剪接因子突变,GoT-Splice提供了一个强大的解决方案.
- 这一进步有助于更深入地了解各种生物环境中的拼接失调.
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