期刊
NUCLEIC ACIDS RESEARCH
卷 42, 期 16, 页码 -出版社
OXFORD UNIV PRESS
DOI: 10.1093/nar/gku606
关键词
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资金
- NSF [DBI-1253293]
- NIH [HG007233-01]
- Defense Advanced Research Projects Agency Living Foundries program [HR0011-12-C-0065]
- California Institute for Quantitative Biosciences (QB3)
- Rogers Family Foundation
- UCSF/Sandler Foundation Program for Breakthrough Biomedical Research
- University of California Proof of Concept Program
- Cancer Research Coordinating Committee of the University of California
- NSF
- Direct For Biological Sciences
- Div Of Biological Infrastructure [1253293] Funding Source: National Science Foundation
Cell sorting is a central tool in life science research for analyzing cellular heterogeneity or enriching rare cells out of large populations. Although methods like FACS and FISH-FC can characterize and isolate cells from heterogeneous populations, they are limited by their reliance on antibodies, or the requirement to chemically fix cells. We introduce a new cell sorting technology that robustly sorts based on sequence-specific analysis of cellular nucleic acids. Our approach, PCR-activated cell sorting (PACS), uses TaqMan PCR to detect nucleic acids within single cells and trigger their sorting. With this method, we identified and sorted prostate cancer cells from a heterogeneous population by performing > 132 000 simultaneous single-cell TaqMan RT-PCR reactions targeting vimentin mRNA. Following vimentin-positive droplet sorting and downstream analysis of recovered nucleic acids, we found that cancer-specific genomes and transcripts were significantly enriched. Additionally, we demonstrate that PACS can be used to sort and enrich cells via TaqMan PCR reactions targeting single-copy genomic DNA. PACS provides a general new technical capability that expands the application space of cell sorting by enabling sorting based on cellular information not amenable to existing approaches.
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