An Engineered CRISPR-Cas9 Mouse Line for Simultaneous Readout of Lineage Histories and Gene Expression Profiles in Single Cells.
An Engineered CRISPR-Cas9 Mouse Line for Simultaneous Readout of Lineage Histories and Gene Expression Profiles in Single Cells.
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DOI：
10.1016/j.cell.2020.04.048
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发表时间：
2020-06-11
期刊：
影响因子：
64.5
通讯作者：
Camargo FD
中科院分区：
文献类型：
--
作者：
Bowling S;Sritharan D;Osorio FG;Nguyen M;Cheung P;Rodriguez-Fraticelli A;Patel S;Yuan WC;Fujiwara Y;Li BE;Orkin SH;Hormoz S;Camargo FD
追踪细胞的谱系历史是回答生物学中各种基本问题的关键。细胞祖先信息与其他分子读数的耦合代表了该领域的重要目标。在这里，我们描述了CARLIN（用于CRISPR阵列修复LINESTRA追踪）小鼠系和相应的分析工具，其可用于同时询问体内单细胞的谱系和转录组信息。该模型利用CRISPR技术在发育或成年期间的任何时间点以诱导方式生成多达44，000个转录条形码，与顺序条形码兼容，并且完全由基因定义。我们已经使用CARLIN来识别胎肝造血干细胞（HSC）克隆活性的内在偏差，并揭示了HSC对损伤反应中以前未被认识到的克隆瓶颈。CARLIN还允许无偏地鉴定与HSC活性相关的转录特征，而无需细胞分选。
Tracing the lineage history of cells is key to answering diverse and fundamental questions in biology. Coupling of cell ancestry information with other molecular readouts represents an important goal in the field. Here, we describe the CARLIN (for CRISPR Array Repair LINeage tracing) mouse line and corresponding analysis tools that can be used to simultaneously interrogate the lineage and transcriptomic information of single cells in vivo. This model exploits CRISPR technology to generate up to 44,000 transcribed barcodes in an inducible fashion at any point during development or adulthood, is compatible with sequential barcoding, and is fully genetically defined. We have used CARLIN to identify intrinsic biases in the activity of fetal liver hematopoietic stem cell (HSC) clones and to uncover a previously unappreciated clonal bottleneck in the response of HSCs to injury. CARLIN also allows the unbiased identification of transcriptional signatures associated with HSC activity without cell sorting.
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影响因子：
64.5
作者：
Kretzschmar, Kai;Watt, Fiona M.
通讯作者：
Watt, Fiona M.
影响因子：
64.8
作者：
Busch, Katrin;Klapproth, Kay;Rodewald, Hans-Reimer
通讯作者：
Rodewald, Hans-Reimer
DOI：
10.1126/science.aaf7907
发表时间：
2016-07-29
期刊：
Science (New York, N.Y.)
影响因子：
--
作者：
McKenna A;Findlay GM;Gagnon JA;Horwitz MS;Schier AF;Shendure J
通讯作者：
Shendure J
DOI：
10.1073/pnas.1607774113
发表时间：
2016-11-22
影响因子：
11.1
作者：
Orlitsky, Alon;Suresh, Ananda Theertha;Wu, Yihong
通讯作者：
Wu, Yihong
影响因子：
64.8
作者：
Alemany, Anna;Florescu, Maria;van Oudenaarden, Alexander
通讯作者：
van Oudenaarden, Alexander