ChIP-seq protocol for sperm cells and embryos to assess environmental impacts and epigenetic inheritance

STAR Protoc. 2021 Jun 11;2(2):100602. doi: 10.1016/j.xpro.2021.100602. eCollection 2021 Jun 18.

Abstract

In the field of epigenetic inheritance, delineating molecular mechanisms implicated in the transfer of paternal environmental conditions to descendants has been elusive. This protocol details how to track sperm chromatin intergenerationally. We describe mouse model design to probe chromatin states in single mouse sperm and techniques to assess pre-implantation embryo chromatin and gene expression. We place emphasis on how to obtain high-quality and quantifiable data sets in sperm and embryos, as well as highlight the limitations of working with low input. For complete details on the use and execution of this protocol, please refer to Lismer et al. (2021).

Keywords: Biophysics; ChIPseq; Chromatin immunoprecipitation (ChIP); Gene Expression; Genomics; Model Organisms; RNAseq; Sequencing.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Chromatin / metabolism
  • Chromatin Immunoprecipitation / methods*
  • Embryo, Mammalian / drug effects*
  • Embryo, Mammalian / metabolism
  • Epigenesis, Genetic*
  • Female
  • Humans
  • Male
  • Mice
  • RNA, Messenger / genetics
  • Sequence Analysis, RNA / methods
  • Spermatozoa / drug effects*
  • Spermatozoa / metabolism

Substances

  • Chromatin
  • RNA, Messenger

Grants and funding