CITE-Seq (cellular indexing of transcriptomes and epitopes) is a sequencing-based method that simultaneously quantifies cell surface protein and transcriptomic data within a single cell readout. Studying cells concurrently at transcriptomic and proteomic levels can offer unprecedented insights into new cell types, disease states, or other conditions.
Before the development of CITE-Seq, studying the transcriptome and protein expression simultaneously was challenging to achieve at the single-cell level. Using transcriptional information to understand cell physiology is helpful, but it doesn't provide the entire picture as cell function is incomplete without protein expression information. RNA analysis cannot accurately measure post-transcriptional and translational modifications such as protein degradation, isoform detection, and glycosylation. A long-standing method to study surface protein expression, such as flow cytometry, is robust but has been traditionally limited by the number of targets that can be analyzed, plus it cannot simultaneously provide transcriptomic data. CITE-seq accomplishes both of these goals in a single assay to provide a comprehensive view of single-cell function.
CITE-Seq solves the problem of detecting a limited number of proteins while using single-cell sequencing to study transcripts in an unbiased way. Wielding the technology of barcoded antibodies, CITE-Seq is capable of analyzing a near-limitless number of cell markers along with transcripts in a single run. Combined with existing single-cell sequencing methods, our sequencing platforms ensure a smooth workflow to identify novel advancements with high throughput, scalability, and speed.
CITE-seq uses unique oligo-tagged antibodies to identify surface proteins, using sequencing as a readout. The number of barcodes that can be conjugated to antibodies surpasses the number of fluorophores or heavy metal tags used in flow cytometry or CyTOF, expanding the number of proteins that can be measured simultaneously with RNA. These barcoded antibodies are part of a unique workflow that produces protein and nucleic acid data using next-generation sequencing (NGS) technologies.
CITE-Seq has enabled important discoveries by using multiplexed quantification of proteins and transcriptomic data all within single cells. Here are just a few examples of CITE-Seq's capabilities:
RNA expression and protein sequencing (REAP-seq) and CITE-seq are based on similar principles. With the development of commercial reagents to support both protocols, many researchers now refer to them interchangeably or use the terms proteogenomics or multiomic analysis.
BD AbSeq Antibody-Oligonucleotide Conjugates allow for the detection of proteins and mRNA expression in a single experiment. These reagents are designed to work seamlessly with the BD Rhapsody Single-Cell Analysis system.
BioLegend offers multiple formats of antibodies for the simultaneous detection of protein and RNA or DNA by sequencing using their TotalSeq oligo-conjugated antibodies for single-cell analysis.
10x Genomics offers a wide range of solutions for the co-detection of protein and gene expression in single cells.
Read how researchers mapped 197 surface markers to cellular profiles using BD Rhapsody and Illumina NGS systems at single-cell resolution in a single experiment.
Optimize BEN-Seq using Illumina NGS and BioLegend TotalSeq Antibodies for bulk protein and gene expression profiling in Th1 cells.
Learn how to optimize TotalSeq-A (BioLegend) antibodies along with RNASeq to add a cost-effective proteomic analysis into your workflow.
Simultaneously profile gene and protein expression for B-cells and T-cells at single-cell resolution using 10x Genomics Chromium Single Cell Immune Profiling and Illumina sequencing systems.
Widely available user-friendly tools provide a simple yet powerful way for you to analyze data without a bioinformatics background. 10x Genomics, BD, and Biolegend offer robust analytics pipelines and visualization tools to aid the interpretation of CITE-Seq data. Alternatively, you can conveniently normalize and integrate gene and protein expression data from CITE-Seq experiments using free open source software such as Seurat or CiteFuse.
Combine gene and protein expression assays into a single workflow.
Quantify data using a single sequencing run.
Discover insights using powerful analysis and visualization software platforms.
Experts from S2 Genomics, Dolomite Bio, and Illumina will provide key insights and tips to optimize your single-cell research for scRNA-seq, SNuc-seq, ATAC-seq, and more.
Whether you are new or experienced in single cell sequencing techniques, this webinar will provide you with the best up-to-date practices for sample preparation and quality control for your workflow.
Learn valuable tips on single-cell sequencing to gain higher discovery power for your research.
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