In Situ Chromatin-Binding Assay Using Epifluorescent Microscopy in S. pombe
Chromatin-associated proteins play critical roles in many cellular processes, including gene expression, epigenetic regulation, DNA repair, recombination, and replication. Especially, epigenetic landscape, shaped by a variety of chromatin-binding proteins, is dynamic and regulated in a context-dependent manner. In situ chromatin-binding assay is a powerful but simple tool to investigate how proteins, such as epigenetic components, associate with chromatin. This approach relies on the fact that chromatin bound proteins are more resistant to detergent extraction. Here, we describe a protocol for the in situ chromatin-binding assay used in Schizosaccaromyces pombe.
Key wordsEpigenetics In situ chromatin binding H3 CENP-A GFP Fission yeast Schizosaccaromyces pombe
We thank Dr. Qianhua Dong, Hyoju Ban, and Dr. David Aristizabal Corrales for reading the manuscript. This work was supported by National Institutes of Health grant R01GM106037 (to F.L.) and NSF grant MCB-1330557 (to F.L.). F. L. is a Pew Scholar in the Biomedical Sciences, supported by The Pew Charitable Trusts.
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