In this episode of the Epigenetics Podcast, we caught up with Active Motif scientists Casidee McDonough from Epigenetic Services and Kyle Tanguay from R&D to talk about technical details of the CUT&Tag protocol and current developments around this method in our R&D Team.
CUT&Tag, which is short for Cleavage Under Targets and Tagmentation, is a molecular biology method that is used to investigate interactions between proteins and DNA and to identify DNA binding sites for their protein of interest. Although CUT&Tag is similar in some ways to ChIP assays, the starting material for CUT&Tag is live, permeabilized cells or isolated cell nuclei, rather than cells or tissue that have been crosslinked with formaldehyde as is the case when performing ChIP. The CUT&Tag method is very sensitive and has been reported to work with as few as 60 cells for some histone modifications. The ability to work with such small numbers of cells is an advantage for researchers working on specific cell types, such as rare neuronal populations, pancreatic islets, or stem cells that are difficult to obtain in large numbers.
In this Episode we discuss the CUT&Tag workflow in detail, talk about the challenges and pitfalls, give guidelines on how to do a good CUT&Tag experiment and offer a glimpse into the future of CUT&Tag product development at Active Motif.
References
Comprehensive Guide to Understanding and Using CUT&Tag Assays
Library QC for ATAC-Seq and CUT&Tag | AKA “Does My Library Look Okay?”
Kaya-Okur, H.S., Wu, S.J., Codomo, C.A. et al. CUT&Tag for efficient epigenomic profiling of small samples and single cells. Nat Commun 10, 1930 (2019). https://doi.org/10.1038/s41467-019-09982-5
Podcast: Chromatin Profiling: From ChIP to CUT&RUN, CUT&Tag and CUTAC (Steven Henikoff)
CUT&Tag-validated antibodies
Contact
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Email: podcast@activemotif.com
Enhancer-Promoter Interactions During Development (Yad Ghavi-Helm)
The Effect of Vitamin D on the Epigenome (Folami Ideraabdullah)
H3K4me3, Set-Proteins, Isw1 and their Role in Transcription (Jane Mellor)
The role of DNA Methylation in Epilepsy (Katja Kobow)
MacroH2A Function in Development and Disease (Emily Bernstein)
Spatio-Temporal Alterations in Chromosome Dynamics (Jane Skok)
Chromatin Organization During Development and Disease (Marieke Oudelaar)
Enhancers and Chromatin Remodeling in Mammary Gland Development (Camila dos Santos)
The Role of SMCHD1 in Development and Disease (Marnie Blewitt)
Single-Molecule Imaging of the Epigenome (Efrat Shema)
Heterochromatin Protein 1 and its Influence on the Structure of Chromatin (Serena Sanulli)
The Effects of Early Life Stress on Mammalian Development (Catherine J. Peña)
Targeting COMPASS to Cure Childhood Leukemia (Ali Shilatifard)
Effects of Non-Enzymatic Covalent Histone Modifications on Chromatin (Yael David)
ATAC-Seq, scATAC-Seq and Chromatin Dynamics in Single-Cells (Jason Buenrostro)
Synthetic Chromatin Epigenetics (Karmella Haynes)
Development of Integrative Machine Learning Tools for Neurodegenerative Diseases (Enrico Glaab)
Ultraconserved Enhancers and Enhancer Redundancy (Diane Dickel)
Variants of Core Histones: Modulators of Chromatin Structure and Function (Sandra Hake)
DNA Methylation and Mammalian Development (Déborah Bourc'his)
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