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Simultaneous multiplexed amplicon sequencing and transcriptome profiling in single cells.

ABSTRACT: We describe droplet-assisted RNA targeting by single-cell sequencing (DART-seq), a versatile technology that enables multiplexed amplicon sequencing and transcriptome profiling in single cells. We applied DART-seq to simultaneously characterize the non-A-tailed transcripts of a segmented dsRNA virus and the transcriptome of the infected cell. In addition, we used DART-seq to simultaneously determine the natively paired, variable region heavy and light chain amplicons and the transcriptome of B lymphocytes.

SUBMITTER: Saikia M

PROVIDER: S-EPMC6378878 | biostudies-literature | 2019 Jan

REPOSITORIES: biostudies-literature

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Simultaneous multiplexed amplicon sequencing and transcriptome profiling in single cells.

Saikia Mridusmita M Burnham Philip P Keshavjee Sara H SH Wang Michael F Z MFZ Heyang Michael M Moral-Lopez Pablo P Hinchman Meleana M MM Danko Charles G CG Parker John S L JSL De Vlaminck Iwijn I

Nature methods 20181217 1

We describe droplet-assisted RNA targeting by single-cell sequencing (DART-seq), a versatile technology that enables multiplexed amplicon sequencing and transcriptome profiling in single cells. We applied DART-seq to simultaneously characterize the non-A-tailed transcripts of a segmented dsRNA virus and the transcriptome of the infected cell. In addition, we used DART-seq to simultaneously determine the natively paired, variable region heavy and light chain amplicons and the transcriptome of B l ...[more]

PMID: 30559431

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Simultaneous multiplexed amplicon sequencing and transcriptome profiling in single cells

Project description:We developed a tunable ligation technology to alter beads used for to capture mRNA for single-cell RNA sequencing (e.g. Drop-seq); we call this technique droplet assisted RNA targeting by single cell sequencing, or DART-seq. In the study, we applied the new technology to capture viral genome transcripts of recombinant type 3 Dearing orthoreovirus infecting murine L929 cells. In addition to capture of targeted viral gene transcripts, we were also able to simultaneosly pull-down polyadenylated mRNA from the L929 cells to observe host-pathogen interactions. We also applied the technology to efficiently measure natively paired heavy and light chain amplicons from B lymphocyte cells in a collection of human peripheral blood mononuclear cells.

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