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
The Impact of Sequence Variation on Transcription Factor Binding (Sven Heinz)
Comparing CUT&Tag to ENCODE ChIP-Seq in Alzheimer's Disease Samples (Sarah Marzi)
The Role of Hat1p in Chromatin Assembly (Mark Parthun)
The Impact of Paternal Diet on Offspring Metabolism (Upasna Sharma)
H3K36me3, H4K16ac and Cryptic Transcription in Ageing (Weiwei Dang)
Split-Pool Recognition of Interactions by Tag Extension (SPRITE) (Mitch Guttman)
MLL Proteins in Mixed-Lineage Leukemia (Yali Dou)
Sex-biased Imprinting and DNA Regulatory Landscapes During Reprogramming (Sam Buckberry)
BET Proteins and Their Role in Chromosome Folding and Compartmentalization (Kyle Eagen)
Epigenetic Underpinnings of Human Addiction (Francesca Telese & Jessica Zhou)
H3K79 Methylation, DOT1L, and FOXG1 in Neural Development (Tanja Vogel)
Function of Insulators in 3D Genome Folding (Maria Gambetta)
Contribution of the Estrogen Receptor to Breast Cancer Progression (Jason Carroll)
Inheritance of Transcriptional Memory by Mitotic Bookmarking (Sheila Teves)
Differential Methylated Regions in Autism Spectrum Disorders (Janine La Salle)
DNA Damage in Longevity and Ageing (Björn Schumacher)
The Impact of Chromatin Modifiers on Disease Development and Progression (Capucine van Rechem)
Long-Range Transcriptional Control by 3D Chromosome Structure (Luca Giorgetti)
Transgenerational Inheritance and Epigenetic Imprinting in Plants (Mary Gehring)
When is a Peak a Peak? (Claudio Cantù)
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