An Engineered CRISPR-Cas9 Mouse Line for Simultaneous Readout of Lineage Histories and Gene Expression Profiles in Single Cells

被引：188

作者：

Bowling, Sarah ^{[1
,2
]}

Sritharan, Duluxan ^{[3
,4
]}

Osorio, Fernando G. ^{[1
,2
]}

Nguyen, Maximilian ^{[4
,5
]}

Cheung, Priscilla ^{[1
,2
]}

Rodriguez-Fraticelli, Alejo ^{[1
,2
]}

Patel, Sachin ^{[1
,2
]}

Yuan, Wei-Chien ^{[1
,2
]}

Fujiwara, Yuko ^{[6
]}

Li, Bin E. ^{[6
,7
]}

Orkin, Stuart H. ^{[6
,8
]}

Hormoz, Sahand ^{[4
,5
,9
]}

Camargo, Fernando D. ^{[1
,2
]}

机构：

[1] Boston Childrens Hosp, Stem Cell Program, Boston, MA 02115 USA

[2] Harvard Univ, Dept Stem Cell & Regenerat Biol, Cambridge, MA 02138 USA

[3] Harvard Univ, Harvard Grad Program Biophys, Cambridge, MA 02138 USA

[4] Dana Farber Canc Inst, Dept Data Sci, Boston, MA 02115 USA

[5] Harvard Med Sch, Dept Syst Biol, Boston, MA 02115 USA

[6] Harvard Med Sch, Div Hematol Oncol, Boston Childrens Hosp, Boston, MA 02115 USA

[7] Dana Farber Canc Inst, Dept Pediat Oncol, Boston, MA 02115 USA

[8] Howard Hughes Med Inst, Boston, MA 02115 USA

[9] Broad Inst MIT & Harvard, Cambridge, MA 02142 USA

来源：

CELL | 2020年 / 181卷 / 06期

基金：

加拿大自然科学与工程研究理事会;

关键词：

HEMATOPOIETIC STEM-CELLS; IDENTIFICATION; HOMEOSTASIS; EFFICIENT; MICE;

D O I：

10.1016/j.cell.2020.04.048

中图分类号：

Q5 [生物化学]; Q7 [分子生物学];

学科分类号：

071010 ; 081704 ;

摘要：

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 CRISPR array repair lineage tracing (CARLIN) 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. Wehave 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.

引用

页码：1410 / +

页数：40

共 48 条

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