Simultaneous Quantification of Single-Cell Proteomes and Transcriptomes in Integrated Fluidic Circuits.

Mandi Wong, Carol Kosman, Liane Takahashi,Naveen Ramalingam

Methods in molecular biology (Clifton, N.J.)(2022)

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摘要
Understanding the principles of gene regulation at single-cell resolution would require measurement and integration of multiple methods such as DNA mutation profiling, open chromatin profiling, RNA expression, protein quantification, and DNA methylation. Recent developments in single-cell multi-omic technologies have enabled integration of these modes in various combinations.With the advent of RNA expression and protein sequencing assay (REAP-seq), researchers can simultaneously analyze protein and gene expression within the same cell. In REAP-seq , cells are labeled with antibodies conjugated to unique DNA sequences. A barcode of 8 nucleotides can allow up to 65,536 unique barcodes for multiplex analysis of proteins, circumventing the limitations of fluorescence (~17 targets). Here, we describe the implementation of REAP-seq assay in the Fluidigm C1™ mRNA Seq HT (high-throughput) v2 system.
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关键词
Gene expression,Genomics,Immunology,Lymphocytes,Methods,Multi-omics,Protein expression,REAP-seq,RNA,Sequencing,Single cell analysis
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