Mapping the chromatin accessibility landscape of zebrafish embryogenesis at single-cell resolution by SPATAC-seq

Affiliations
  • 1School of Medicine, Tsinghua University, Beijing, China.
  • 2Peking University-Tsinghua University-National Institute of Biological Sciences Joint Graduate Program, Tsinghua University, Beijing, China.
  • 3School of Life Sciences, Tsinghua University, Beijing, China.
  • 4Tsinghua University-Peking University Center for Life Sciences, Beijing, China.
  • 5School of Life Sciences, Tsinghua University, Beijing, China. mengam@mail.tsinghua.edu.cn.
  • 6Tsinghua University-Peking University Center for Life Sciences, Beijing, China. mengam@mail.tsinghua.edu.cn.
  • 7School of Medicine, Tsinghua University, Beijing, China. xlan@tsinghua.edu.cn.
  • 8Peking University-Tsinghua University-National Institute of Biological Sciences Joint Graduate Program, Tsinghua University, Beijing, China. xlan@tsinghua.edu.cn.
  • 9Tsinghua University-Peking University Center for Life Sciences, Beijing, China. xlan@tsinghua.edu.cn.
  • 10Ministry of Education Key Laboratory of Bioinformatics, Tsinghua University, Beijing, China. xlan@tsinghua.edu.cn.

Published on:

Abstract

Currently, the dynamic accessible elements that determine regulatory programs responsible for the unique identity and function of each cell type during zebrafish embryogenesis lack detailed study. Here we present SPATAC-seq: a split-pool ligation-based assay for transposase-accessible chromatin using sequencing. Using SPATAC-seq, we profiled chromatin accessibility in more than 800,000 individual nuclei across 20 developmental stages spanning the sphere stage to the early larval protruding mouth stage. Using this chromatin accessibility map, we identified 604 cell states and inferred their developmental relationships. We also identified 959,040 candidate cis-regulatory elements (cCREs) and delineated development-specific cCREs, as well as transcription factors defining diverse cell identities. Importantly, enhancer reporter assays confirmed that the majority of tested cCREs exhibited robust enhanced green fluorescent protein expression in restricted cell types or tissues. Finally, we explored gene regulatory programs that drive pigment and notochord cell differentiation. Our work provides a valuable open resource for exploring driver regulators of cell fate decisions in zebrafish embryogenesis.