期刊
NATURE BIOTECHNOLOGY
卷 40, 期 3, 页码 374-+出版社
NATURE PORTFOLIO
DOI: 10.1038/s41587-021-01065-5
关键词
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资金
- NHGRI [5RM1 HG006193]
- EMBO Long-Term fellowship
- Stanford Science Fellowship
- Emmy Noether fellowship of the German Research Foundation [LU 2336/2-1]
- Ludwig Family Foundation
- Howard Hughes Medical Institute
The study introduces a novel method called PHAGE-ATAC for simultaneous single-cell measurements of protein levels and chromatin accessibility profiles, using mitochondrial DNA-based clonal tracing. PHAGE-ATAC is utilized for multimodal analysis in primary human immune cells, sample multiplexing, intracellular protein analysis, and detection of SARS-CoV-2 spike protein. Additionally, a synthetic high-complexity phage library is constructed for selection of antigen-specific nanobodies, enabling protein detection, cell characterization, and screening with single-cell genomics.
Multimodal measurements of single-cell profiles are proving increasingly useful for characterizing cell states and regulatory mechanisms. In the present study, we developed PHAGE-ATAC (Assay for Transposase-Accessible Chromatin), a massively parallel droplet-based method that uses phage displaying, engineered, camelid single-domain antibodies ('nanobodies') for simultaneous single-cell measurements of protein levels and chromatin accessibility profiles, and mitochondrial DNA-based clonal tracing. We use PHAGE-ATAC for multimodal analysis in primary human immune cells, sample multiplexing, intracellular protein analysis and the detection of SARS-CoV-2 spike protein in human cell populations. Finally, we construct a synthetic high-complexity phage library for selection of antigen-specific nanobodies that bind cells of particular molecular profiles, opening an avenue for protein detection, cell characterization and screening with single-cell genomics. Protein epitopes and chromatin accessibility are measured in single cells using phage display.
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