Abstract
Recently, frequent somatic mutations at histone genes have been detected in high grade pediatric brain tumor, chondroblastoma, and giant cell tumor of bone. These mutant histones are also termed oncohistones. Since oncohistone proteins co-exist with wild type histone proteins in cells, it is critically important to understand how they promote tumorigenesis. Here, we describe two methods to analyze the impact of these oncohistones on histone modification and epigenome, including the expression of oncohistone from a transgene and the utilization of CRISPR/Cas9 system to knock-in specific oncohistone mutations. The methods described are useful for the initial characterization of oncohistones. Other methods such as ChIP-seq and RNA-seq, which analyze the effect of oncohistone mutations genome wide, are not detailed in this protocol.
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Fang, D., Wang, H., Zhang, Z. (2018). Probing the Function of Oncohistones Using Mutant Transgenes and Knock-In Mutations. In: Orsi, G., Almouzni, G. (eds) Histone Variants. Methods in Molecular Biology, vol 1832. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-8663-7_19
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DOI: https://doi.org/10.1007/978-1-4939-8663-7_19
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