Identification of potential nuclear reprogramming and differentiation factors by a novel selection method for cloning chromatin-binding proteins.
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.
Wang
Liu
L
Department of Cell Biology and Genetics, College of Life Science, Peking University, Beijing, PR China.
Zheng
Aihua
A
Yi
Ling
L
Xu
Chongren
C
Ding
Mingxiao
M
Deng
Hongkui
H
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
D002843:Chromatin; D004268:DNA-Binding Proteins; D019151:Peptide Library