Assay of Genome-Wide Transcriptome and Secreted Proteins on the Same Single Immune Cells by Microfluidics and RNA Sequencing.
Given vast heterogeneity of immune cells, searching for gene expression and transcriptional networks belonging to specific cellular functions such as cytokine production has been challenging. To overcome this limitation, we developed a splittable single-cell microchip that integrates a high-density antibody array for cytokine protein detection, while the same single cells with protein profiles can be subsequently sequenced to obtain the genome-wide transcriptome. Combined with bioinformatics algorithms, we discovered a subgroup of highly coexpressed genes correlating with TNFα secretion in mouse macrophage cells. This technology and the data analysis may lead to an unprecedented understanding of regulation mechanisms of the immune system and have the potential to impact disease treatment and drug discovery.
Multiplex Biotechnology Laboratory, Department of Chemistry, University at Albany, State University of New York , Albany, New York 12222, United States.
Justin
J
George
Multiplex Biotechnology Laboratory, Department of Chemistry, University at Albany, State University of New York , Albany, New York 12222, United States.|Cancer Research Center, University at Albany, State University of New York , Rensselaer, New York 12144, United States.
Jun
J
Wang
D000818:Animals; D004797:Enzyme-Linked Immunosorbent Assay; D005456:Fluorescent Dyes / Q000737:chemistry; D016678:Genome; D059014:High-Throughput Nucleotide Sequencing; D016753:Interleukin-10 / Q000032:analysis; D057230:Limit of Detection; D051379:Mice; D008807:Mice, Inbred BALB C; D044085:Microfluidics; D008856:Microscopy, Fluorescence; D009000:Monocytes / Q000166:cytology / Q000378:metabolism; D000067996:RAW 264.7 Cells; D012333:RNA, Messenger / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism; D017423:Sequence Analysis, RNA; D059010:Single-Cell Analysis; D059467:Transcriptome; D014409:Tumor Necrosis Factor-alpha / Q000032:analysis*
D005456:Fluorescent Dyes; D012333:RNA, Messenger; D014409:Tumor Necrosis Factor-alpha; D016753:Interleukin-10