中国科学院脑科学与智能技术卓越中心Yuejun Chen和中国科学院上海营养与健康研究所Wu Wei共同合作，近期取得重要工作进展。他们研究利用CRISPR条形码技术全面绘制小鼠脑内的单细胞时空谱系。相关研究成果2023年7月17日在线发表于《自然—方法学》杂志上。

据介绍，发育神经科学的一个基本兴趣在于绘制大脑内完整单细胞谱系的能力。

研究人员开发了一种基于CRISPR编辑的谱系特异性追踪（CREST）方法，用于Cre小鼠的克隆追踪。然后，研究人员使用了两种基于CREST的互补策略来绘制发育中的小鼠腹侧中脑（vMB）的单细胞谱系。通过应用snapshotting CREST（snapCREST），研究人员构建了一个发展中vMB的时空谱系概况，并确定了六个祖细胞原型，这些原型可以代表单个vMB祖细胞的主要克隆命运，以及底板中指定谷氨酸能、多巴胺能或两种神经元的三个不同的克隆谱系。研究人员进一步创建了pandaCREST（祖细胞和衍生物相关CREST），以使体内祖细胞的转录组与其分化潜能相关联。研究人员确定了多巴胺能神经元的多种起源，并证明转录组定义的祖细胞类型包括异质祖细胞，每个祖细胞都具有不同的克隆命运和分子特征。

因此，CREST方法和策略允许进行全面的单细胞谱系分析，这可以为神经规范背后的分子程序提供新的见解。

附：英文原文

Title: Comprehensive spatiotemporal mapping of single-cell lineages in developing mouse brain by CRISPR-based barcoding

Author: Xie, Lianshun, Liu, Hengxin, You, Zhiwen, Wang, Luyue, Li, Yiwen, Zhang, Xinyue, Ji, Xiaoshan, He, Hui, Yuan, Tingli, Zheng, Wenping, Wu, Ziyan, Xiong, Man, Wei, Wu, Chen, Yuejun

Issue&Volume: 2023-07-17

Abstract: A fundamental interest in developmental neuroscience lies in the ability to map the complete single-cell lineages within the brain. To this end, we developed a CRISPR editing-based lineage-specific tracing (CREST) method for clonal tracing in Cre mice. We then used two complementary strategies based on CREST to map single-cell lineages in developing mouse ventral midbrain (vMB). By applying snapshotting CREST (snapCREST), we constructed a spatiotemporal lineage landscape of developing vMB and identified six progenitor archetypes that could represent the principal clonal fates of individual vMB progenitors and three distinct clonal lineages in the floor plate that specified glutamatergic, dopaminergic or both neurons. We further created pandaCREST (progenitor and derivative associating CREST) to associate the transcriptomes of progenitor cells in vivo with their differentiation potentials. We identified multiple origins of dopaminergic neurons and demonstrated that a transcriptome-defined progenitor type comprises heterogeneous progenitors, each with distinct clonal fates and molecular signatures. Therefore, the CREST method and strategies allow comprehensive single-cell lineage analysis that could offer new insights into the molecular programs underlying neural specification.

DOI: 10.1038/s41592-023-01947-3

Source: https://www.nature.com/articles/s41592-023-01947-3