美国波士顿儿童医院Fernando D. Camargo团队开发出一个具有高克隆条形码多样性的小鼠模型，用于单细胞的联合谱系、转录组学和表观基因组学分析。相关论文于2023年10月17日发表在《细胞》杂志上。

他们开发了DARLIN，一种利用末端脱氧核苷酸转移酶(TdT)和30个CRISPR靶点的诱导型Cas9条形码小鼠系。DARLIN是可诱导的，在组织中产生大量的谱系条形码，并且能够在约70%的谱化单细胞中检测编辑过的条形码。使用DARLIN，他们检测了发育中的造血干细胞(HSC)的命运偏差，并揭示了HSC迁移的独特特征。

此外，他们用DARLIN建立了一个联合转录组学和表观基因组学单细胞测量方案，发现细胞克隆记忆与全基因组DNA甲基化有关，而不是与基因表达或染色质可及性有关。DARLIN将使谱系关系及其分子特征在不同组织和生理背景下的高分辨率研究成为可能。

研究人员表示，细胞谱系历史及其分子状态编码了组织发育和体内平衡的基本原理。目前的谱系记录小鼠模型缺乏条形码多样性和单细胞谱系覆盖，无法分析由数百万细胞组成的组织。

附：英文原文

Title: A mouse model with high clonal barcode diversity for joint lineage, transcriptomic, and epigenomic profiling in single cells

Author: Li Li, Sarah Bowling, Sean E. McGeary, Qi Yu, Bianca Lemke, Karel Alcedo, Yuemeng Jia, Xugeng Liu, Mark Ferreira, Allon M. Klein, Shou-Wen Wang, Fernando D. Camargo

Issue&Volume: 2023-10-17

Abstract: Cellular lineage histories and their molecular states encode fundamental principlesof tissue development and homeostasis. Current lineage-recording mouse models haveinsufficient barcode diversity and single-cell lineage coverage for profiling tissuescomposed of millions of cells. Here, we developed DARLIN, an inducible Cas9 barcodingmouse line that utilizes terminal deoxynucleotidyl transferase (TdT) and 30 CRISPRtarget sites. DARLIN is inducible, generates massive lineage barcodes across tissues,and enables the detection of edited barcodes in ～70% of profiled single cells. UsingDARLIN, we examined fate bias within developing hematopoietic stem cells (HSCs) andrevealed unique features of HSC migration. Additionally, we established a protocolfor joint transcriptomic and epigenomic single-cell measurements with DARLIN and foundthat cellular clonal memory is associated with genome-wide DNA methylation ratherthan gene expression or chromatin accessibility. DARLIN will enable the high-resolutionstudy of lineage relationships and their molecular signatures in diverse tissues andphysiological contexts.

DOI: 10.1016/j.cell.2023.09.019

Source: https://www.cell.com/cell/fulltext/S0092-8674(23)01040-1

期刊信息

Cell：《细胞》，创刊于1974年。隶属于细胞出版社，最新IF：66.85

官方网址：https://www.cell.com/

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