Protocol to profile snATAC-seq datasets and motif enrichment analysis during zebrafish early embryogenesis

STAR Protoc. 2024 Dec 20;5(4):103501. doi: 10.1016/j.xpro.2024.103501. Epub 2024 Dec 12.

Abstract

The scarcity of zebrafish-specific motif databases presents a challenge to the analysis of transcription factor (TF) motif within zebrafish single-cell assay for transposase-accessible chromatin using sequencing (scATAC-seq) data, thus hindering the identification of regulatory elements throughout zebrafish embryonic development. Here, we provide a protocol to analyze single-nucleus chromatin accessibility dataset during zebrafish early embryogenesis. We describe steps for fragment file retrieval, sample integration, quality control, Latent Semantic Indexing (LSI) clustering, and peak calling via ArchR. Crucially, we detail procedures for zebrafish-specific motif database construction, motif enrichment, and TF footprinting analysis. For complete details on the use and execution of this protocol, please refer to Lin et al.1.

Keywords: bioinformatics; developmental biology; model organisms; single cell.

MeSH terms

  • Animals
  • Chromatin / chemistry
  • Chromatin / genetics
  • Chromatin / metabolism
  • Embryo, Nonmammalian / metabolism
  • Embryonic Development* / genetics
  • Nucleotide Motifs / genetics
  • Single-Cell Analysis / methods
  • Transcription Factors / genetics
  • Transcription Factors / metabolism
  • Zebrafish* / embryology
  • Zebrafish* / genetics

Substances

  • Transcription Factors
  • Chromatin