title: Identification of potential nuclear reprogramming and differentiation factors by a novel selection method for cloning chromatin-binding proteins.
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.
authors.affiliation: Department of Cell Biology and Genetics, College of Life Science, Peking University, Beijing, PR China.
authors.forename: Liu
authors.initials: L
authors.lastname: Wang
authors.forename: Aihua
authors.initials: A
authors.lastname: Zheng
authors.forename: Ling
authors.initials: L
authors.lastname: Yi
authors.forename: Chongren
authors.initials: C
authors.lastname: Xu
authors.forename: Mingxiao
authors.initials: M
authors.lastname: Ding
authors.forename: Hongkui
authors.initials: H
authors.lastname: Deng
mesh_terms: D000818:Animals; D017123:Bacteriophage T7 / Q000235:genetics / Q000378:metabolism; D001681:Biological Assay / Q000379:methods*; D002454:Cell Differentiation / Q000502:physiology*; D002467:Cell Nucleus / Q000378:metabolism*; D002843:Chromatin; D047369:Chromatin Immunoprecipitation; D004268:DNA-Binding Proteins / Q000378:metabolism*; D004622:Embryo, Mammalian / Q000033:anatomy & histology; D005347:Fibroblasts / Q000166:cytology / Q000502:physiology; D015723:Gene Library; D051379:Mice; D008969:Molecular Sequence Data; D019151:Peptide Library; D011897:Random Allocation; D013234:Stem Cells / Q000166:cytology / Q000502:physiology
chemical_list: D002843:Chromatin; D004268:DNA-Binding Proteins; D019151:Peptide Library