Effective ribosomal RNA depletion for single-cell total RNA-seq by scDASH

PeerJ

article

Danson S.C. Loi¹  Lei Yu¹  Angela R. Wu¹ 

[1] Division of Life Science, Hong Kong University of Science and Technology;Department of Chemical and Biological Engineering, Hong Kong University of Science and Technology;Hong Kong Branch of Guangdong Southern Marine Science and Engineering Laboratory ,(Guangzhou), Hong Kong University of Science and Technology

关键词: Single-cell transcriptomics;    scRNA-seq;    rRNA depletion;    CRISPR;   

DOI  :  10.7717/peerj.10717

学科分类：社会科学、人文和艺术（综合）

来源: Inra

【 摘 要 】

A decade since its invention, single-cell RNA sequencing (scRNA-seq) has become a mainstay technology for profiling transcriptional heterogeneity in individual cells. Yet, most existing scRNA-seq methods capture only polyadenylated mRNA to avoid the cost of sequencing non-messenger transcripts, such as ribosomal RNA (rRNA), that are usually not of-interest. Hence, there are not very many protocols that enable single-cell analysis of total RNA. We adapted a method called DASH (Depletion of Abundant Sequences by Hybridisation) to make it suitable for depleting rRNA sequences from single-cell total RNA-seq libraries. Our analyses show that our single-cell DASH (scDASH) method can effectively deplete rRNAs from sequencing libraries with minimal off-target non-specificity. Importantly, as a result of depleting the rRNA, the rest of the transcriptome is significantly enriched for detection.

【 授权许可】

CC BY