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通过组织中确定性条形码进行高空间分辨率多态序列化

Yang Liu¹, Mingyu Yang¹, Yanxiang Deng¹

¹Department of Biomedical Engineering, Yale University, New Haven, CT 06520, USA; Yale Stem Cell Center and Yale Cancer Center, Yale School of Medicine, New Haven, CT 06520, USA.

|November 14, 2020

概括

组织中的确定性条形编码用于空间奥米克测序 (DBiT-seq) 可以同时映射固定组织中的mRNA和蛋白质. 这种技术为各种生物研究应用提供高分辨率的空间数据.

科学领域:

空间奥米克
分子生物学
基因组学

背景情况:

空间奥米克技术对于理解组织架构和细胞异质性至关重要.
在高分辨率下同时绘制多个生物分子 (例如mRNA和蛋白质) 仍然是一个挑战.

研究的目的:

开发一种新的方法,即组织中的确定性条形码用于空间奥米克测序 (DBiT-seq),用于固定组织幻灯片中的mRNA和蛋白质的共同映射.
通过下一代测序 (NGS) 实现高分辨率的空间转录和蛋白质组.

主要方法:

DBiT-seq使用并行微流体通道将DNA条形码传递到组织滑块表面.
两个条形码集 (A和B) 的顺序传递和现场绑定为每个组织像素创建独特的二维条形码.
使用下一代测序 (NGS) 来读取空间数据.

主要成果:

在甲固定小鼠胚胎组织中,DBiT-seq成功地共映射了mRNA和蛋白质.
高分辨率地图显示器官生成过程中的主要组织类型以及微血管和有色表皮等细结构.
来自10μm像素的基因表达特征与单细胞转录组聚集在一起,使得细胞类型和空间分布的快速识别.

结论:

DBiT-seq提供了一个易于使用的空间学方法,可供没有微流体专业知识的研究人员使用.
这种技术在发育生物学,癌症生物学,神经科学和临床病理学方面具有广泛的应用.
DBiT-seq通过在现场进行综合多态分析来推进空间生物学领域.

关键词:

高空间分辨率在现场编码小鼠胚胎下一代测序空间多态学

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