TAS-Seq is a robust and sensitive amplification method for bead-based scRNA-seq

Commun Biol. 2022 Jun 27;5(1):602. doi: 10.1038/s42003-022-03536-0.

Abstract

Single-cell RNA-sequencing (scRNA-seq) is valuable for analyzing cellular heterogeneity. Cell composition accuracy is critical for analyzing cell-cell interaction networks from scRNA-seq data. However, droplet- and plate-based scRNA-seq techniques have cell sampling bias that could affect the cell composition of scRNA-seq datasets. Here we developed terminator-assisted solid-phase cDNA amplification and sequencing (TAS-Seq) for scRNA-seq based on a terminator, terminal transferase, and nanowell/bead-based scRNA-seq platform. TAS-Seq showed high tolerance to variations in the terminal transferase reaction, which complicate the handling of existing terminal transferase-based scRNA-seq methods. In murine and human lung samples, TAS-Seq yielded scRNA-seq data that were highly correlated with flow-cytometric data, showing higher gene-detection sensitivity and more robust detection of important cell-cell interactions and expression of growth factors/interleukins in cell subsets than 10X Chromium v2 and Smart-seq2. Expanding TAS-Seq application will improve understanding and atlas construction of lung biology at the single-cell level.

MeSH terms

  • Animals
  • DNA, Complementary / genetics
  • Gene Expression Profiling* / methods
  • Humans
  • Mice
  • Sequence Analysis, RNA / methods
  • Single-Cell Analysis* / methods
  • Transferases

Substances

  • DNA, Complementary
  • Transferases