Single-Cell Antibody Sequencing in Atherosclerosis Research

The transcriptomic information obtained by single cell RNA sequencing (scRNA-seq) can be supplemented by information on the cell surface phenotype by using oligonucleotide-tagged monoclonal antibodies (scAb-Seq). This is of particular importance in immune cells, where the correlation between mRNA an...

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Bibliographic Details
Published in:Methods in molecular biology (Clifton, N.J.) Vol. 2419; p. 765
Main Authors: Saigusa, Ryosuke, Durant, Christopher P, Suryawanshi, Vasantika, Ley, Klaus
Format: Journal Article
Language:English
Published: United States 2022
Subjects:
Animals
Atherosclerosis - genetics
Gene Expression Profiling
High-Throughput Nucleotide Sequencing
Mice
Sequence Analysis, RNA
Single-Cell Analysis
Transcriptome
RNA expression and protein sequencing assay (REAP-seq)
TotalSeq
Surface markers
Cellular indexing of transcriptomes and epitopes by sequencing (CITE-seq)
AbSeq
ISSN:1940-6029, 1940-6029
Online Access:Get more information
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Summary:The transcriptomic information obtained by single cell RNA sequencing (scRNA-seq) can be supplemented by information on the cell surface phenotype by using oligonucleotide-tagged monoclonal antibodies (scAb-Seq). This is of particular importance in immune cells, where the correlation between mRNA and cell surface expression is very weak. scAb-Seq is facilitated by the availability of commercial antibodies and antibody mixes. Now panels of up to 200 antibodies are available for human and mouse cells. Proteins are detected by antibodies conjugated to a tripartite DNA sequence that contains a primer for amplification and sequencing, a unique oligonucleotide that acts as an antibody barcode and a poly(dA) sequence, simultaneously detecting extension of antibody-specific DNA sequences and cDNAs in the same poly(dT)-primed reaction. For each cell, surface protein expression is captured and sequenced along with the cell's transcriptome. Here, we list the steps needed to produce antibody sequencing data from tissue or blood cells.
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ISSN:1940-6029
1940-6029
DOI:10.1007/978-1-0716-1924-7_46