Targeting individual cells by barcode in pooled sequence libraries.

Nucleic Acids Res
Authors
Keywords
Abstract

Transcriptional profiling of thousands of single cells in parallel by RNA-seq is now routine. However, due to reliance on pooled library preparation, targeting analysis to particular cells of interest is difficult. Here, we present a multiplexed PCR method for targeted sequencing of select cells from pooled single-cell sequence libraries. We demonstrated this molecular enrichment method on multiple cell types within pooled single-cell RNA-seq libraries produced from primary human blood cells. We show how molecular enrichment can be combined with FACS to efficiently target ultra-rare cell types, such as the recently identified AXL+SIGLEC6+ dendritic cell (AS DC) subset, in order to reduce the required sequencing effort to profile single cells by 100-fold. Our results demonstrate that DNA barcodes identifying cells within pooled sequencing libraries can be used as targets to enrich for specific molecules of interest, for example reads from a set of target cells.

Year of Publication
2019
Journal
Nucleic Acids Res
Volume
47
Issue
1
Pages
e4
Date Published
2019 01 10
ISSN
1362-4962
DOI
10.1093/nar/gky856
PubMed ID
30256981
PubMed Central ID
PMC6326790
Links
Grant list
RM1 HG006193 / HG / NHGRI NIH HHS / United States
U24 AI118668 / AI / NIAID NIH HHS / United States