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
Active Motif on Twitter
Epigenetics Podcast on Twitter
Active Motif on LinkedIn
Active Motif on Facebook
Email: podcast@activemotif.com
Chromatin Analysis using Mass Spectrometry (Axel Imhof)
Chromatin Profiling: From ChIP to CUT&RUN, CUT&Tag and CUTAC (Steven Henikoff)
Effects of DNA Methylation on Diabetes (Charlotte Ling)
Nutriepigenetics: The Effects of Diet on Behavior (Monica Dus)
Investigating the Dynamics of Epigenetic Plasticity in Cancer with Single Cell Technologies (Céline Vallot)
Epigenetic Regulation of Stem Cell Self-Renewal and Differentiation (Margaret “Peggy” Goodell)
Genome-Wide Investigation of Epigenetic Marks and Nucleosome Positioning (Keji Zhao)
The Role of lncRNAs in Tumor Growth and Treatment (Sarah Diermeier)
Unraveling Mechanisms of Chromosome Formation (Job Dekker)
Transcription and Polycomb in Inheritance and Disease (Danny Reinberg)
The Epigenetics of COVID-19
Epigenetic Reprogramming During Mammalian Development (Wolf Reik)
In vivo Nucleosome Structure and Dynamics (Srinivas Ramachandran)
Pioneer Transcription Factors and Their Influence on Chromatin Structure (Ken Zaret)
The Role of Small RNAs in Transgenerational Inheritance in C. elegans (Oded Rechavi)
Development of Site-Specific ChIP Technologies (Hodaka Fujii)
Regulation of Chromatin Organization by Histone Chaperones (Geneviève Almouzni)
How the "Fragile Nucleosome" Science Community Came to Life (Christine Cucinotta, Melvin Noe Gonzalez)
Epigenetic Influence on Memory Formation and Inheritance (Isabelle Mansuy)
Influence of Dynamic RNA Methylation on Gene Expression (Chuan He)
Create your
podcast in
minutes
It is Free
DNA Today: A Genetics Podcast
The Psychic Elephant Radio Podcast
Museum of the Missing
Sasquatch Chronicles
Hidden Brain