Purification of Zygotically Transcribed RNA through Metabolic Labeling of Early Zebrafish Embryos

Methods Mol Biol. 2017:1605:121-131. doi: 10.1007/978-1-4939-6988-3_8.

Abstract

Early embryonic development in all known metazoans is characterized by a transcriptionally silent phase, during which development is under control of maternally loaded protein and RNA. The zygotic genome becomes transcriptionally active after a series of rapid reductive cleavage divisions. In this chapter, we present a method to metabolically label, purify, and analyze newly transcribed RNAs in early zebrafish embryos. We previously used this method, which is adaptable to other embryos and systems, to determine the onset of zygotic transcription activation and identify the first zygotic transcripts.

Keywords: 4-sUTP; Biotinylation; RNA; Transcription; Zebrafish; Zygotic genome activation.

MeSH terms

  • Animals
  • Embryonic Development
  • Gene Expression Regulation, Developmental
  • RNA, Messenger / analysis*
  • RNA, Messenger / chemistry
  • Transcriptional Activation
  • Uridine Triphosphate / administration & dosage
  • Uridine Triphosphate / analogs & derivatives*
  • Zebrafish / embryology*
  • Zebrafish / genetics
  • Zebrafish Proteins / genetics*

Substances

  • RNA, Messenger
  • Zebrafish Proteins
  • 4-thiouridine triphosphate
  • Uridine Triphosphate