Targeting individual cells by barcode in pooled sequence libraries.
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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
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Journal | Nucleic Acids Res
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Volume | 47
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Issue | 1
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Pages | e4
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Date Published | 2019 01 10
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ISSN | 1362-4962
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DOI | 10.1093/nar/gky856
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PubMed ID | 30256981
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PubMed Central ID | PMC6326790
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Grant list | RM1 HG006193 / HG / NHGRI NIH HHS / United States
U24 AI118668 / AI / NIAID NIH HHS / United States
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