Second-Strand Synthesis-Based Massively Parallel scRNA-Seq Reveals Cellular States and Molecular Features of Human Inflammatory Skin Pathologies
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1-s2.0-S107476132030409X-main.pdf
Description
Published version
Size
7.02 MB
Format
Adobe PDF
Checksum (MD5)
8802c1586664ccb2c8a3daf16527f3cf
Author(s)
Hughes, Travis K
Wadsworth, Marc H
Gierahn, Todd M
Do, Tran
Weiss, David
Andrade, Priscila R
Ma, Feiyang
de Andrade Silva, Bruno J
Shao, Shuai
Tsoi, Lam C
Date Issued
2020
Journal
Immunity
Publisher
Elsevier BV
Version
Final published version
Abstract
© 2020 The Authors High-throughput single-cell RNA-sequencing (scRNA-seq) methodologies enable characterization of complex biological samples by increasing the number of cells that can be profiled contemporaneously. Nevertheless, these approaches recover less information per cell than low-throughput strategies. To accurately report the expression of key phenotypic features of cells, scRNA-seq platforms are needed that are both high fidelity and high throughput. To address this need, we created Seq-Well S3 (“Second-Strand Synthesis”), a massively parallel scRNA-seq protocol that uses a randomly primed second-strand synthesis to recover complementary DNA (cDNA) molecules that were successfully reverse transcribed but to which a second oligonucleotide handle, necessary for subsequent whole transcriptome amplification, was not appended due to inefficient template switching. Seq-Well S3 increased the efficiency of transcript capture and gene detection compared with that of previous iterations by up to 10- and 5-fold, respectively. We used Seq-Well S3 to chart the transcriptional landscape of five human inflammatory skin diseases, thus providing a resource for the further study of human skin inflammation.
MIT Department
Massachusetts Institute of Technology. Institute for Medical Engineering & Science
Ragon Institute of MGH, MIT and Harvard
Koch Institute for Integrative Cancer Research at MIT
Massachusetts Institute of Technology. Department of Chemical Engineering
Massachusetts Institute of Technology. Department of Chemistry
Terms of Use
Creative Commons Attribution 4.0 International license
Persistent DSpace Link
DOI of Published Version
10.1016/J.IMMUNI.2020.09.015
https://doi.org/10.1016/J.IMMUNI.2020.09.015
