使用条形码衰变谱系追踪研究非遗传编码状态的遗传和传播的协议
Yelyzaveta Shlyakhtina1, Bianca Bloechl1, Katherine L Moran2
1Cell Plasticity & Epigenetics Lab, Cancer Research UK Manchester Institute, The University of Manchester, Manchester M20 4BX, UK; Cell Plasticity & Epigenetics Lab, Cancer Research UK - Cancer Research UK Scotland Institute, The University of Glasgow, Glasgow G61 1BD, UK.
STAR protocols
|January 5, 2024
概括
我们开发了条形码衰变谱系追踪,随后进行单细胞转录组分析 (BdLT-Seq),以追踪克隆进化. 这种方法可以比较同源细胞系之间的分子特征.
科学领域:
- 分子生物学分子生物学
- 基因组学就是基因组学.
- 细胞生物学 细胞生物学
背景情况:
- 了解克隆进化在各种生物过程中至关重要.
- 现有的谱系追踪方法可能缺乏所需的分辨率或定向追踪.
- 同源细胞种群提供了一个可控的系统来研究非遗传变异.
研究的目的:
- 提出一个新的条形码衰变谱系追踪协议,结合单细胞转录组分析 (BdLT-Seq).
- 为了使克隆进化的定向评估,同时保持细胞同质性.
- 为了促进基因相同的细胞系之间的分子特征的比较.
主要方法:
- BdLT-Seq. 的实验设计.
- 构建和转换条形码的插曲记者.
- 条码检索,序列库准备和样本复杂化.
- 单细胞转录组分析和数据解释.
主要成果:
- BdLT-Seq协议提供了一个全面的谱系追踪框架.
- 该方法允许对细胞群的定向跟踪.
- 保持同质性使分子表型的直接比较成为可能.
结论:
- BdLT-Seq是研究克隆动态和异质性的强大技术.
- 该协议允许研究同源细胞系中的非遗传差异.
- 描述的方法促进了细胞进化和分化的研究.
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