Simultaneous Quantification of Spatial Genome Positioning and Transcriptomics in Single Cells with scDam&T-Seq
Lochs, Silke J.A.; Kind, Jop
(2022) Sexton, Tom (ed.), Spatial Genome Organization, pp. 215 - 241
Methods in Molecular Biology, pp. 215 - 241
(Part of book)
Abstract
Spatial genome organization is considered to play an important role in mammalian cells, by guiding gene expression programs and supporting lineage specification. Yet it is still an outstanding question in the field what the direct impact of spatial genome organization on gene expression is. To elucidate this relationship further, we
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have recently developed scDam&T-seq, a method that simultaneously quantifies protein–DNA interactions and transcriptomes in single cells. This method efficiently combines two preexisting methods: DamID for measuring protein–DNA contacts and CEL-Seq2 for quantification of the transcriptome in single cells. scDam&T-seq has been successfully applied to measure DNA contacts with the nuclear lamina, while at the same time revealing the effect of these contacts on gene expression. This method is applicable to many different proteins of interest and can thereby aid in studying the relationship between protein–DNA interactions and gene expression in single cells.
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Keywords: CEL-Seq2, DamID, Lamina-associated domains, Protein–DNA interactions, Single-cell genomics, Single-Cell Analysis/methods, Animals, Transcriptome, Mammals/genetics, Genome, Proteins/genetics, DNA/genetics, Genetics, Molecular Biology, Research Support, Non-U.S. Gov't, Journal Article
ISSN: 1064-3745
ISBN: 978-1-0716-2496-8
978-1-0716-2499-9
978-1-0716-2497-5
Publisher: Humana New York, Humana New York
Note: Funding Information: We would like to thank the members of the Kind lab for their critical reading of the manuscript and their helpful suggestions and comments. Work in the Kind lab on the scDam&T-seq technique was funded by the European Research Council Starting grant (no. ERC-StG 678423-EpiID) and a Dutch Research Council (NWO) Open grant (no. 824.15.019). J.K. and S.J.A.L. are funded by an NWO ALW/VIDI grant (no. 161.339). The Oncode Institute is supported by the KWF Dutch Cancer Society. Publisher Copyright: © 2022, The Author(s), under exclusive license to Springer Science+Business Media, LLC, part of Springer Nature.
(Peer reviewed)