Multiplexed Analysis of Retinal Gene Expression and Chromatin Accessibility using scRNA-Seq and scATAC-Seq

J Vis Exp. 2021 Mar 12:(169):10.3791/62239. doi: 10.3791/62239.

Abstract

Powerful next generation sequencing techniques offer robust and comprehensive analysis to investigate how retinal gene regulatory networks function during development and in disease states. Single-cell RNA sequencing allows us to comprehensively profile gene expression changes observed in retinal development and disease at a cellular level, while single-cell ATAC-Seq allows analysis of chromatin accessibility and transcription factor binding to be profiled at similar resolution. Here the use of these techniques in the developing retina is described, and MULTI-Seq is demonstrated, where individual samples are labeled with a modified oligonucleotide-lipid complex, enabling researchers to both increase the scope of individual experiments and substantially reduce costs.

Publication types

  • Video-Audio Media

MeSH terms

  • Chromatin / chemistry
  • Chromatin / genetics
  • Chromatin / metabolism*
  • Chromatin Immunoprecipitation Sequencing / methods*
  • High-Throughput Nucleotide Sequencing / methods*
  • Humans
  • RNA, Small Cytoplasmic / analysis
  • RNA, Small Cytoplasmic / genetics
  • RNA, Small Cytoplasmic / metabolism*
  • Retina / metabolism*
  • Sequence Analysis, RNA / methods*
  • Single-Cell Analysis / methods*

Substances

  • Chromatin
  • RNA, Small Cytoplasmic