Identification of potential nuclear reprogramming and differentiation factors by a novel selection method for cloning chromatin-binding proteins

Biochem Biophys Res Commun. 2004 Dec 3;325(1):302-7. doi: 10.1016/j.bbrc.2004.10.028.

Abstract

Nuclear reprogramming is critical for animal cloning and stem cell creation through nuclear transfer, which requires extensive remodeling of chromosomal architecture involving dramatic changes in chromatin-binding proteins. To understand the mechanism of nuclear reprogramming, it is critical to identify chromatin-binding factors specify the reprogramming process. In this report, we have developed a high-throughput selection method, based on T7 phage display and chromatin immunoprecipitation, to isolate chromatin-binding factors expressed in mouse embryonic stem cells using primary mouse embryonic fibroblast chromatin. Seven chromatin-binding proteins have been isolated by this method. We have also isolated several chromatin-binding proteins involved in hepatocyte differentiation. Our method provides a powerful tool to rapidly and selectively identify chromatin-binding proteins. The method can be used to study epigenetic modification of chromatin during nuclear reprogramming, cell differentiation, and transdifferentiation.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Bacteriophage T7 / genetics
  • Bacteriophage T7 / metabolism
  • Biological Assay / methods*
  • Cell Differentiation / physiology*
  • Cell Nucleus / metabolism*
  • Chromatin
  • Chromatin Immunoprecipitation
  • DNA-Binding Proteins / metabolism*
  • Embryo, Mammalian / anatomy & histology
  • Fibroblasts / cytology
  • Fibroblasts / physiology
  • Gene Library
  • Mice
  • Molecular Sequence Data
  • Peptide Library
  • Random Allocation
  • Stem Cells / cytology
  • Stem Cells / physiology

Substances

  • Chromatin
  • DNA-Binding Proteins
  • Peptide Library