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A Scalable Strand-Specific Protocol Enabling Full-Length Total RNA Sequencing From Single Cells.

Simon Haile¹, Richard D Corbett¹, Veronique G LeBlanc¹

¹Canada's Michael Smith Genome Sciences Centre, BC Cancer, Vancouver, BC, Canada.

Frontiers in Genetics

|June 21, 2021

Summary

This study introduces a new single-cell RNA sequencing (scRNAseq) method for high-throughput, full-length total RNA profiling. This advanced scRNAseq technique overcomes limitations of existing methods, enabling detailed analysis of cellular heterogeneity.

Keywords:

RNAseq cellenONE full-length single-cell total RNA

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Area of Science:

Genomics
Molecular Biology
Bioinformatics

Background:

Bulk RNA sequencing (RNAseq) provides gene expression data from cell pools.
Single-cell RNA sequencing (scRNAseq) enables high-resolution analysis of cellular heterogeneity.
Existing scRNAseq methods have limitations, including restricted transcript coverage (3'/5' termini) and limited throughput.

Purpose of the Study:

To develop a novel scRNAseq protocol addressing current technical constraints.
To enable high-throughput, full-length, total RNA sequencing at the single-cell level.
To profile both polyadenylated and non-polyadenylated transcripts comprehensively.

Main Methods:

Development of a novel single-cell RNA sequencing protocol.
High-throughput sequencing of full-length, total RNA from single cells.
Strand-specific sequencing for detailed transcript analysis.

Main Results:

The new protocol sequences full-length, total RNA at single-cell resolution.
It generates strand-specific data for both polyadenylated and non-polyadenylated transcripts.
The method allows profiling beyond transcript termini and identifies cell types in heterogeneous samples.

Conclusions:

The developed scRNAseq method overcomes limitations of existing protocols.
It provides comprehensive transcriptomic data at single-cell resolution.
This advancement facilitates deeper understanding of cellular heterogeneity in complex biological systems like tumors.