Epigenetics is a key regulatory layer determining how the universal blueprint – the genome – is utilized within each cell and during disease.
Students will perform an epigenetic assay, which involves detecting the localization of histone modifications using CUT&Tag (Kaya-Okur, Nat Commun, 2019 doi.org/10.1038/s41467-019-09982-5), and the binding of transcription factors and other DNA binding proteins via the CUT&RUN assay (Skene, eLife, 2017, doi.org/10.7554/eLife.21856).
CUT&Tag can be utilized to determine the localisation of multiple proteins on a single-cell level with engineered enzymes and adapter oligos (Stuart, Nat Biotechnol, 2022, doi.org/10.1038/s41587-022-01588-5 and Bartosovic, Nat Biotechnol, 2022, doi.org/10.1038/s41587-022-01535-4). Additionally, we try to acquire transcriptomic data from the same cell to integrate with single-cell RNA-sequencing results.
Students will learn various methods, including PCR, RTqPCR, cell culture, antibody staining, sequencing library preparation, preprocessing, and analysis of CUT&RUN and CUT&Tag data.