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Chromatin Profiling: From ChIP to CUT&RUN, CUT&Tag and CUTAC (Steven Henikoff)
저장한 시리즈 ("피드 비활성화" status)
When? This feed was archived on September 02, 2022 22:36 (). Last successful fetch was on July 28, 2022 16:40 ()
Why? 피드 비활성화 status. 잠시 서버에 문제가 발생해 팟캐스트를 불러오지 못합니다.
What now? You might be able to find a more up-to-date version using the search function. This series will no longer be checked for updates. If you believe this to be in error, please check if the publisher's feed link below is valid and contact support to request the feed be restored or if you have any other concerns about this.
Manage episode 290022079 series 2369335
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.
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.
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.
References
Takehito Furuyama, Steven Henikoff (2009) Centromeric nucleosomes induce positive DNA supercoils (Cell) DOI: 10.1016/j.cell.2009.04.049
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: 10.1073/pnas.1118898109
Jorja G. Henikoff, Jitendra Thakur, … Steven Henikoff (2015) A unique chromatin complex occupies young α-satellite arrays of human centromeres (Science Advances) DOI: 10.1126/sciadv.1400234
Epigenomics Methods: ChEC-Seq, CUT&RUN, AutoCUT&RUN, Improved CUT&RUN, CUT&Tag, Automated CUT&Tag, CUTAC, scCUT&Tag.
Related Episodes
The Role of Non-Histone Proteins in Chromosome Structure and Function During Mitosis (Bill Earnshaw)
Hi-C and Three-Dimensional Genome Sequencing (Erez Lieberman Aiden)
In Vivo Nucleosome Structure and Dynamics (Srinivas Ramachandran)
Contact
80 에피소드
저장한 시리즈 ("피드 비활성화" status)
When? This feed was archived on September 02, 2022 22:36 (). Last successful fetch was on July 28, 2022 16:40 ()
Why? 피드 비활성화 status. 잠시 서버에 문제가 발생해 팟캐스트를 불러오지 못합니다.
What now? You might be able to find a more up-to-date version using the search function. This series will no longer be checked for updates. If you believe this to be in error, please check if the publisher's feed link below is valid and contact support to request the feed be restored or if you have any other concerns about this.
Manage episode 290022079 series 2369335
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.
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.
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.
References
Takehito Furuyama, Steven Henikoff (2009) Centromeric nucleosomes induce positive DNA supercoils (Cell) DOI: 10.1016/j.cell.2009.04.049
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: 10.1073/pnas.1118898109
Jorja G. Henikoff, Jitendra Thakur, … Steven Henikoff (2015) A unique chromatin complex occupies young α-satellite arrays of human centromeres (Science Advances) DOI: 10.1126/sciadv.1400234
Epigenomics Methods: ChEC-Seq, CUT&RUN, AutoCUT&RUN, Improved CUT&RUN, CUT&Tag, Automated CUT&Tag, CUTAC, scCUT&Tag.
Related Episodes
The Role of Non-Histone Proteins in Chromosome Structure and Function During Mitosis (Bill Earnshaw)
Hi-C and Three-Dimensional Genome Sequencing (Erez Lieberman Aiden)
In Vivo Nucleosome Structure and Dynamics (Srinivas Ramachandran)
Contact
80 에피소드
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