Chromatin Profiling: From ChIP to CUT&RUN, CUT&Tag and CUTAC (Steven Henikoff)

<p class="md-end-block md-p">In this episode of the Epigenetics Podcast, we caught up with Steven Henikoff from the Fred Hutchinson Cancer Research Center in Seattle to talk about his work on Chromatin Profiling: From ChIP to CUT&RUN, CUT&Tag and CUTAC.</p> <p class="md-end-block md-p">In the last few years Steven Henikoff has been developing methods which profile the chromatin landscape by using enzyme tethering. The quest first started with ChEC-Seq, which improved on Uli Laemmli's method of Chromatin endogenous cleavage (ChEC) but used sequencing as a read-out rather than southern blotting. Next, Cleavage Under Targets & Release Using Nuclease (CUT&RUN) was developed by making a fusion protein of Protein A and micrococcal nuclease (MNase), making it possible to achieve antibody-targeted cleavage of chromatin fragments. And finally, Cleavage Under Targets & Tagmenation (CUT&Tag) was developed by using Transposase Tn5 instead of MNase, which adds sequencing adapters and fragments chromatin at the same time, streamlining the protocol even further.</p> <p class="md-end-block md-p">In this episode we discuss how working on centromeres set the stage for Steven Henikoff’s subsequent work, how he developed CUT&RUN and CUT&Tag, what the advantages and disadvantages of those methods are and how he developed all those experiments at home in his garage. </p> <p class="md-end-block md-p"> </p> <p class="md-end-block md-p">References</p> <ul class="ul-list"><li class="md-list-item"> <p class="md-end-block md-p">Takehito Furuyama, Steven Henikoff (2009) Centromeric nucleosomes induce positive DNA supercoils (Cell) DOI: <a href='http://dx.doi.org/10.1016/j.cell.2009.04.049'>10.1016/j.cell.2009.04.049</a></p> </li> <li class="md-list-item"> <p class="md-end-block md-p">Kristina Krassovsky, Jorja G. Henikoff, Steven Henikoff (2012) Tripartite organization of centromeric chromatin in budding yeast (Proceedings of the National Academy of Sciences of the United States of America) DOI: <a href='http://dx.doi.org/10.10