Concept

Single-cell transcriptomics

Résumé
Single-cell transcriptomics examines the gene expression level of individual cells in a given population by simultaneously measuring the RNA concentration (conventionally only messenger RNA (mRNA)) of hundreds to thousands of genes. Single-cell transcriptomics makes it possible to unravel heterogeneous cell populations, reconstruct cellular developmental pathways, and model transcriptional dynamics — all previously masked in bulk RNA sequencing. The development of high-throughput RNA sequencing (RNA-seq) and microarrays has made gene expression analysis a routine. RNA analysis was previously limited to tracing individual transcripts by Northern blots or quantitative PCR. Higher throughput and speed allow researchers to frequently characterize the expression profiles of populations of thousands of cells. The data from bulk assays has led to identifying genes differentially expressed in distinct cell populations, and biomarker discovery. These studies are limited as they provide measurements for whole tissues and, as a result, show an average expression profile for all the constituent cells. This has a couple of drawbacks. Firstly, different cell types within the same tissue can have distinct roles in multicellular organisms. They often form subpopulations with unique transcriptional profiles. Correlations in the gene expression of the subpopulations can often be missed due to the lack of subpopulation identification. Secondly, bulk assays fail to recognize whether a change in the expression profile is due to a change in regulation or composition — for example if one cell type arises to dominate the population. Lastly, when your goal is to study cellular progression through differentiation, average expression profiles can only order cells by time rather than by developmental stage. Consequently, they cannot show trends in gene expression levels specific to certain stages. Recent advances in biotechnology allow the measurement of gene expression in hundreds to thousands of individual cells simultaneously.
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