STRTseq

STRTseq, also known as Single-cell Tagged Reverse Transcription sequencing, is a high-throughput method for profiling the transcriptome of individual cells. It allows for the simultaneous measurement of gene expression and other cellular features. The core principle of STRTseq involves capturing messenger RNA (mRNA) molecules from single cells and tagging them with a unique molecular identifier (UMI) and a cell barcode during the reverse transcription process. This tagging is crucial for distinguishing between multiple copies of the same mRNA molecule and for assigning each transcript to its originating cell.

The STRTseq protocol typically begins with cell lysis and the addition of reagents that facilitate the capture and reverse transcription of mRNA. The UMIs help to correct for amplification bias during downstream sequencing, providing a more accurate representation of the original mRNA abundance. The cell barcodes ensure that reads from different cells are correctly demultiplexed. Following library preparation, the tagged cDNA libraries are sequenced using next-generation sequencing technologies. The resulting data can then be analyzed to quantify gene expression levels in thousands of individual cells, enabling researchers to identify distinct cell populations, study cellular heterogeneity, and investigate developmental processes or disease mechanisms at a single-cell resolution. Variations of STRTseq have been developed to integrate other modalities, such as protein expression, with transcriptomic data.