All multicellular organisms are composed of cells which contain the same DNA. How is it then, that the cells in our stomach look and behave differently from the cells in our brain? The answer lies within the way cells differentiate and select different sets of genes to transcribe, with which to make functional molecules such as RNA and protein.
RNA sequencing is a tool which surveys all the mRNA within cells. It takes advantage of the conserved structure of eukaryotic mRNA, namely the 3’ poly-A tail, and captures all the mRNA within the sample using a common poly-T primer sequence. One of the key benefits of this technique, unlike techniques such as antibody staining which generally target a few specific epitopes that are decided a priori by the researcher, is that it provides an unbiased look of the entire mRNA expression profile, or transcriptome, of the sample.
Single-cell assays have been utilized to reveal insights about cellular heterogeneity which would have been masked in bulk assays. The evolution of RNA sequencing to single cell resolution was first pioneered by Tang and others in 2009 (1) and used mouth pipetting to isolate single cells for analysis. The throughput of the assay has been a key area of innovation within the past decade, with microfluidics based single-cell isolation now able to process thousands of cells per experiment (2, 3).
Single Cell Sequencing Applications
Single-cell RNA sequencing (scRNA-seq) has been leveraged for many applications by researchers. For instance, it has enabled the identification of rare cell types that have unique gene expression profiles. These innovations have enabled detailed annotation of every cell type in the human body, such as the ongoing Human Cell Atlas Project. scRNA-seq has also been used to reconstruct developmental lineages by placing cells on a tree-like-structure depending on their gene expression signature. In addition, new biomarkers have been discovered thanks to scRNA-seq, giving clinicians more options for treating diseases such as cancer that have a high degree of inter and intra-patient heterogeneity.
Fluent BioSciences PIPseq V Single Cell 3′ RNA Kit
Fluent BioSciences’ mission is to make single-cell assays more accessible for all researchers scRNA-seq still has many barriers to entry including high consumables cost, need for specialized capital instrumentation, and significant expertise to process and analyze samples . With our PIPseq V SIngle Cell 3′ RNA Kits, we eliminate these barriers to entry and provide a sensitive, flexible, and scalable solution to expand accessibility and application of these valuable methods.
We are excited to engage with customers interested in applying Fluent’s uniquely enabling scRNAseq solution, and have demonstrated the application of our approach in a wide and growing range of applications and tissue types including drug treatment of cultured cells, pathogen-host interactions, enriched organoid preparations, and whole-tissue profiling, including breast and brain tissues.
Contact us to discuss the right fit for your single cell project at sales@fluentbio.com.
References
- Tang, F., Barbacioru, C., Wang, Y. et al. mRNA-Seq whole-transcriptome analysis of a single cell. Nature Methods 6, 377–382 (2009)
- Macosko, E. Z., Basu, A., Satija, R. et al.. Highly parallel genome-wide expression profiling of individual cells using nanoliter droplets. Cell 161, 1202-1214 (2015).
- Klein, A. M., Mazutis, L., Akartuna, I. et al.. Droplet barcoding for single-cell transcriptomics applied to embryonic stem cells. Cell 161, 1187-1201 (2015).