Purification of GFP-tagged nuclei from frozen livers of INTACT mice for RNA- and ATAC-sequencing

STAR Protoc. 2021 Sep 8;2(3):100805. doi: 10.1016/j.xpro.2021.100805. eCollection 2021 Sep 17.

Abstract

Isolation of nuclei tagged in specific cell types (INTACT) allows for stress-free and high-throughput analyses of cellular subpopulations. Here, we present an improved protocol for isolation of pure and high-quality GFP-labeled nuclei from frozen livers of INTACT mice, as well as protocols for downstream sequencing analyses. The adaptation to frozen tissue provides a pause point that allows sampling at multiple time points and/or phenotypic characterization of livers prior to nuclei isolation and downstream analyses. For complete details on the use of this protocol, please refer to Loft et al. (2021).

Keywords: Cell Biology; Cell isolation; Cell separation/fractionation; Genomics; Molecular Biology; RNAseq; Sequencing.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Cell Nucleus / chemistry*
  • Cell Nucleus / metabolism
  • Chromatin Immunoprecipitation Sequencing*
  • Cytological Techniques / methods*
  • Female
  • Green Fluorescent Proteins / chemistry*
  • Green Fluorescent Proteins / genetics
  • Green Fluorescent Proteins / metabolism
  • Liver / chemistry
  • Liver / cytology*
  • Male
  • Mice
  • RNA-Seq*

Substances

  • Green Fluorescent Proteins