Use of Chemicals to Inhibit DNA Replication, Transcription, and Protein Synthesis to Study Zygotic Genome Activation

Methods Mol Biol. 2017:1605:191-205. doi: 10.1007/978-1-4939-6988-3_13.

Abstract

Maternal-to-zygotic transition is an event that developmental control of early embryos is switched from oocyte-derived factors to the zygotic genome. Ability to inhibit DNA replication, transcription, and translation is an important tool in studying events, such as zygotic genome activation, during embyogenesis. Here, we describe approaches to block DNA replication, transcription, and translation using chemical inhibitors. Then we also demonstrate how the transcript level of a maternally inherited gene, ten-eleven translocation methylcytosine dioxygenase 3, responses to the chemical treatments.

Keywords: Cycloheximide; Oocytes; Pig embryos; TET3; Zygotic genome activation; α-Amanitin.

MeSH terms

  • Alpha-Amanitin / pharmacology*
  • Animals
  • Cycloheximide / pharmacology*
  • DNA Methylation
  • DNA Replication / drug effects
  • Dioxygenases / genetics
  • Maternal Inheritance
  • Nucleic Acid Synthesis Inhibitors / pharmacology*
  • Protein Biosynthesis / drug effects
  • Protein Synthesis Inhibitors / pharmacology*
  • Swine / embryology*
  • Swine / genetics
  • Transcription, Genetic / drug effects
  • Transcriptional Activation

Substances

  • Alpha-Amanitin
  • Nucleic Acid Synthesis Inhibitors
  • Protein Synthesis Inhibitors
  • Cycloheximide
  • Dioxygenases