Abstract
In multicellular organisms, the initiation and maintenance of specific cell types often require the activity of cell type-specific transcriptional regulators. Understanding their roles in gene regulation is crucial but probing their DNA targets in vivo, especially in a genome-wide manner, remains a technical challenge with their limited expression. To improve the sensitivity of chromatin immunoprecipitation (ChIP) for detecting the cell type-specific signals, we have developed the Maximized Objects for Better Enrichment (MOBE)-ChIP, where ChIP is performed at a substantially larger experimental scale and under low background conditions. Here, we describe the procedure in the study of transcription factors in the model plant Arabidopsis. However, with some modifications, the technique should also be implemented in other systems. Besides cell type-specific studies, MOBE-ChIP can also be used as a general strategy to improve ChIP signals.
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Acknowledgments
This work was supported by funds from the National University of Singapore to O.S.L. (AcRF Tier 1 R-154-000-A16-114, and start-up funds R154000695133 and R154000A06651). The authors declare no competing financial interests.
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Wang, S., Lau, O.S. (2018). MOBE-ChIP: Probing Cell Type-Specific Binding Through Large-Scale Chromatin Immunoprecipitation. In: Visa, N., Jordán-Pla, A. (eds) Chromatin Immunoprecipitation. Methods in Molecular Biology, vol 1689. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-7380-4_15
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DOI: https://doi.org/10.1007/978-1-4939-7380-4_15
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Publisher Name: Humana Press, New York, NY
Print ISBN: 978-1-4939-7379-8
Online ISBN: 978-1-4939-7380-4
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