1. Embryonic stem cell–based mapping of developmental transcriptional programs
- Author
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Mazzoni, Esteban O, Mahony, Shaun, Iacovino, Michelina, Morrison, Carolyn A, Mountoufaris, George, Closser, Michael, Whyte, Warren A, Young, Richard A, Kyba, Michael, Gifford, David K, and Wichterle, Hynek
- Subjects
Stem Cell Research ,Human Genome ,Regenerative Medicine ,Stem Cell Research - Nonembryonic - Non-Human ,Genetics ,Biotechnology ,Stem Cell Research - Embryonic - Non-Human ,Stem Cell Research - Embryonic - Human ,Underpinning research ,1.1 Normal biological development and functioning ,Animals ,Cell Differentiation ,Chromatin Immunoprecipitation ,DNA-Binding Proteins ,Embryonic Stem Cells ,Gene Expression Regulation ,Developmental ,Genome-Wide Association Study ,High-Throughput Nucleotide Sequencing ,Mice ,Motor Neurons ,Sequence Analysis ,DNA ,Transcription Factors ,Biological Sciences ,Technology ,Medical and Health Sciences ,Developmental Biology - Abstract
The study of developmentally regulated transcription factors by chromatin immunoprecipitation and deep sequencing (ChIP-seq) faces two major obstacles: availability of ChIP-grade antibodies and access to sufficient number of cells. We describe versatile genome-wide analysis of transcription-factor binding sites by combining directed differentiation of embryonic stem cells and inducible expression of tagged proteins. We demonstrate its utility by mapping DNA-binding sites of transcription factors involved in motor neuron specification.
- Published
- 2011