Single-cell analysis of regions of interest (SCARI) using a photosensitive tag.

Autor: van der Leun AM; Division of Molecular Oncology & Immunology, Oncode Institute, The Netherlands Cancer Institute, Amsterdam, Netherlands., Hoekstra ME; Division of Molecular Oncology & Immunology, Oncode Institute, The Netherlands Cancer Institute, Amsterdam, Netherlands., Reinalda L; Department of Bio-Organic Synthesis, Leiden Institute of Chemistry, Leiden University, Leiden, Netherlands., Scheele CLGJ; Division of Molecular Pathology, Oncode Institute, The Netherlands Cancer Institute, Amsterdam, Netherlands.; VIB-KULeuven Center for Cancer Biology, Leuven, Belgium., Toebes M; Division of Molecular Oncology & Immunology, Oncode Institute, The Netherlands Cancer Institute, Amsterdam, Netherlands., van de Graaff MJ; Department of Bio-Organic Synthesis, Leiden Institute of Chemistry, Leiden University, Leiden, Netherlands.; SeraNovo, Leiden, Netherlands., Chen LYY; Division of Molecular Pathology, Oncode Institute, The Netherlands Cancer Institute, Amsterdam, Netherlands., Li H; Department of Immunology, Weizmann Institute of Science, Rehovot, Israel.; Shenzhen Institute of Synthetic Biology, Shenzhen, China., Bercovich A; Department of Computer Science and Applied Mathematics and Department of Biological Regulation, Weizmann Institute, Rehovot, Israel., Lubling Y; Department of Computer Science and Applied Mathematics and Department of Biological Regulation, Weizmann Institute, Rehovot, Israel.; Cancer Research UK Cambridge Institute, Cambridge, UK., David E; Department of Immunology, Weizmann Institute of Science, Rehovot, Israel., Thommen DS; Division of Molecular Oncology & Immunology, The Netherlands Cancer Institute, Amsterdam, Netherlands., Tanay A; Department of Computer Science and Applied Mathematics and Department of Biological Regulation, Weizmann Institute, Rehovot, Israel., van Rheenen J; Division of Molecular Pathology, Oncode Institute, The Netherlands Cancer Institute, Amsterdam, Netherlands., Amit I; Department of Immunology, Weizmann Institute of Science, Rehovot, Israel., van Kasteren SI; Department of Bio-Organic Synthesis, Leiden Institute of Chemistry, Leiden University, Leiden, Netherlands. s.i.van.kasteren@chem.leidenuniv.nl., Schumacher TN; Division of Molecular Oncology & Immunology, Oncode Institute, The Netherlands Cancer Institute, Amsterdam, Netherlands. t.schumacher@nki.nl.; Department of Hematology, Leiden University Medical Center, Leiden, Netherlands. t.schumacher@nki.nl.
Jazyk: angličtina
Zdroj: Nature chemical biology [Nat Chem Biol] 2021 Nov; Vol. 17 (11), pp. 1139-1147. Date of Electronic Publication: 2021 Sep 09.
DOI: 10.1038/s41589-021-00839-x
Abstrakt: The functional activity and differentiation potential of cells are determined by their interactions with surrounding cells. Approaches that allow unbiased characterization of cell states while at the same time providing spatial information are of major value to assess this environmental influence. However, most current techniques are hampered by a tradeoff between spatial resolution and cell profiling depth. Here, we develop a photocage-based technology that allows isolation and in-depth analysis of live cells from regions of interest in complex ex vivo systems, including primary human tissues. The use of a highly sensitive 4-nitrophenyl(benzofuran) cage coupled to a set of nanobodies allows high-resolution photo-uncaging of different cell types in areas of interest. Single-cell RNA-sequencing of spatially defined CD8 + T cells is used to exemplify the feasibility of identifying location-dependent cell states. The technology described here provides a valuable tool for the analysis of spatially defined cells in diverse biological systems, including clinical samples.
(© 2021. The Author(s), under exclusive licence to Springer Nature America, Inc.)
Databáze: MEDLINE
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