期刊
SCIENCE
卷 371, 期 6528, 页码 481-+出版社
AMER ASSOC ADVANCEMENT SCIENCE
DOI: 10.1126/science.aax2656
关键词
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资金
- IARPA MICrONS [D16PC0008]
- National Institutes of Health [HG005550, HG008525]
- National Science Foundation [DGE1144152]
- Howard Hughes Medical Institute (HHMI) fellowship of the LSRF
- Schmidt Fellows Program at the Broad Institute
- Hertz Foundation Fellowship
- Siebel Scholarship
- NIH Neuroimaging Training Program T32 grant [5T32EB001680]
- Chan Zuckerberg Initiative, an advised fund of Silicon Valley Community Foundation [2017-0525]
- Open Philanthropy Project, Cancer Research UK Grand Challenge grant [C31893/A25050]
- U.S. Army Research Laboratory
- U.S. Army Research Office [W911NF1510548]
- NSF [1734870]
- Chan Zuckerberg Initiative Human Cell Atlas pilot program
- Ludwig Foundation
- HHMI-Simons Faculty Scholars Program
- HHMI investigator program
- National Cancer Institute, National Institutes of Health [HHSN261100039, HHSN261201500003I]
- HFSP long-term fellowship [LT000452/2019-L]
- NIH [UF1NS107697, IDIQ17X149, 1RM1HG008525, 1R01MH103910, 1R01MH114031, 1R01MH110932, 1R01EB024261, 1R01DA045549, 1U19MH114821, 1R01NS102727]
- IARPA [D16PC00008]
- Division Of Undergraduate Education
- Direct For Education and Human Resources [1734870] Funding Source: National Science Foundation
Expansion microscopy is adapted for long-read untargeted and targeted in situ RNA sequencing, enabling highly multiplexed mapping of RNAs from nanoscale to system scale. This technology allows for detailed transcript localization in different cell types and spatial positions, with applications in both animal models and human cancer samples.
Methods for highly multiplexed RNA imaging are limited in spatial resolution and thus in their ability to localize transcripts to nanoscale and subcellular compartments. We adapt expansion microscopy, which physically expands biological specimens, for long-read untargeted and targeted in situ RNA sequencing. We applied untargeted expansion sequencing (ExSeq) to the mouse brain, which yielded the readout of thousands of genes, including splice variants. Targeted ExSeq yielded nanoscale-resolution maps of RNAs throughout dendrites and spines in the neurons of the mouse hippocampus, revealing patterns across multiple cell types, layer-specific cell types across the mouse visual cortex, and the organization and position-dependent states of tumor and immune cells in a human metastatic breast cancer biopsy. Thus, ExSeq enables highly multiplexed mapping of RNAs from nanoscale to system scale.
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