MOBE-ChIP: a large-scale chromatin immunoprecipitation assay for cell type-specific studies

The Plant Journal : for Cell and Molecular Biology
On Sun Lau, Dominique C Bergmann

Abstract

Cell type-specific transcriptional regulators play critical roles in the generation and maintenance of multicellularity. As they are often expressed at low levels, in vivo DNA-binding studies of these regulators by standard chromatin immunoprecipitation (ChIP) assays are technically challenging. We describe here an optimized ChIP protocol termed Maximized Objects for Better Enrichment (MOBE)-ChIP, which enhances the sensitivity of ChIP assays for detecting cell type-specific signals. The protocol, which is based on the disproportional increase of target signals over background at higher scales, uses substantially greater volume of starting materials than conventional ChIPs to achieve high signal enrichment. This technique can capture weak binding events that are ambiguous in standard ChIP assays, and is useful both in gene-specific and whole-genome analysis. This protocol has been optimized for Arabidopsis, but should be applicable to other model systems with minor modifications. The full procedure can be completed within 3 days.

References

Aug 19, 2004·The Plant Journal : for Cell and Molecular Biology·Chris BowlerJerzy Paszkowski
Sep 17, 2005·Nature Methods·Anne-Valérie GendrelVincent Colot
May 15, 2012·The Plant Journal : for Cell and Molecular Biology·Ronny BrandtStephan Wenkel

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Citations

Aug 2, 2016·Trends in Plant Science·Makoto ShirakawaIkuko Hara-Nishimura
Jan 10, 2019·Plant Biotechnology Journal·Shi-Lian QiXue-Jun Hua

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