Improved CUT&RUN chromatin profiling tools
© 2019, Meers et al..
Previously, we described a novel alternative to chromatin immunoprecipitation, CUT&RUN, in which unfixed permeabilized cells are incubated with antibody, followed by binding of a protein A-Micrococcal Nuclease (pA/MNase) fusion protein (Skene and Henikoff, 2017). Here we introduce three enhancements to CUT&RUN: A hybrid protein A-Protein G-MNase construct that expands antibody compatibility and simplifies purification, a modified digestion protocol that inhibits premature release of the nuclease-bound complex, and a calibration strategy based on carry-over of E. coli DNA introduced with the fusion protein. These new features, coupled with the previously described low-cost, high efficiency, high reproducibility and high-throughput capability of CUT&RUN make it the method of choice for routine epigenomic profiling.
| Medienart: |
E-Artikel |
|---|
| Erscheinungsjahr: |
2019 |
|---|---|
| Erschienen: |
2019 |
| Enthalten in: |
Zur Gesamtaufnahme - volume:8 |
|---|---|
| Enthalten in: |
eLife - 8(2019) vom: 24. Juni |
| Sprache: |
Englisch |
|---|
| Beteiligte Personen: |
Meers, Michael P [VerfasserIn] |
|---|
| Links: |
|---|
| Anmerkungen: |
Date Completed 25.02.2020 Date Revised 12.10.2023 published: Electronic GEO: GSE126612 Citation Status MEDLINE |
|---|
| doi: |
10.7554/eLife.46314 |
|---|
| funding: |
|
|---|---|
| Förderinstitution / Projekttitel: |
|
| PPN (Katalog-ID): |
NLM298468859 |
|---|
| LEADER | 01000naa a22002652 4500 | ||
|---|---|---|---|
| 001 | NLM298468859 | ||
| 003 | DE-627 | ||
| 005 | 20231225094227.0 | ||
| 007 | cr uuu---uuuuu | ||
| 008 | 231225s2019 xx |||||o 00| ||eng c | ||
| 024 | 7 | |a 10.7554/eLife.46314 |2 doi | |
| 028 | 5 | 2 | |a pubmed24n0994.xml |
| 035 | |a (DE-627)NLM298468859 | ||
| 035 | |a (NLM)31232687 | ||
| 035 | |a (PII)e46314 | ||
| 040 | |a DE-627 |b ger |c DE-627 |e rakwb | ||
| 041 | |a eng | ||
| 100 | 1 | |a Meers, Michael P |e verfasserin |4 aut | |
| 245 | 1 | 0 | |a Improved CUT&RUN chromatin profiling tools |
| 264 | 1 | |c 2019 | |
| 336 | |a Text |b txt |2 rdacontent | ||
| 337 | |a ƒaComputermedien |b c |2 rdamedia | ||
| 338 | |a ƒa Online-Ressource |b cr |2 rdacarrier | ||
| 500 | |a Date Completed 25.02.2020 | ||
| 500 | |a Date Revised 12.10.2023 | ||
| 500 | |a published: Electronic | ||
| 500 | |a GEO: GSE126612 | ||
| 500 | |a Citation Status MEDLINE | ||
| 520 | |a © 2019, Meers et al. | ||
| 520 | |a Previously, we described a novel alternative to chromatin immunoprecipitation, CUT&RUN, in which unfixed permeabilized cells are incubated with antibody, followed by binding of a protein A-Micrococcal Nuclease (pA/MNase) fusion protein (Skene and Henikoff, 2017). Here we introduce three enhancements to CUT&RUN: A hybrid protein A-Protein G-MNase construct that expands antibody compatibility and simplifies purification, a modified digestion protocol that inhibits premature release of the nuclease-bound complex, and a calibration strategy based on carry-over of E. coli DNA introduced with the fusion protein. These new features, coupled with the previously described low-cost, high efficiency, high reproducibility and high-throughput capability of CUT&RUN make it the method of choice for routine epigenomic profiling | ||
| 650 | 4 | |a Journal Article | |
| 650 | 4 | |a Research Support, N.I.H., Extramural | |
| 650 | 4 | |a Research Support, Non-U.S. Gov't | |
| 650 | 4 | |a chromatin | |
| 650 | 4 | |a chromosomes | |
| 650 | 4 | |a epigenomics | |
| 650 | 4 | |a gene expression | |
| 650 | 4 | |a genetics | |
| 650 | 4 | |a genomics | |
| 650 | 4 | |a human | |
| 650 | 4 | |a spike-in calibration | |
| 650 | 7 | |a Bacterial Proteins |2 NLM | |
| 650 | 7 | |a Chromatin |2 NLM | |
| 650 | 7 | |a IgG Fc-binding protein, Streptococcus |2 NLM | |
| 650 | 7 | |a Recombinant Fusion Proteins |2 NLM | |
| 650 | 7 | |a Staphylococcal Protein A |2 NLM | |
| 650 | 7 | |a Micrococcal Nuclease |2 NLM | |
| 650 | 7 | |a EC 3.1.31.1 |2 NLM | |
| 700 | 1 | |a Bryson, Terri D |e verfasserin |4 aut | |
| 700 | 1 | |a Henikoff, Jorja G |e verfasserin |4 aut | |
| 700 | 1 | |a Henikoff, Steven |e verfasserin |4 aut | |
| 773 | 0 | 8 | |i Enthalten in |t eLife |d 2012 |g 8(2019) vom: 24. Juni |w (DE-627)NLM221831460 |x 2050-084X |7 nnns |
| 773 | 1 | 8 | |g volume:8 |g year:2019 |g day:24 |g month:06 |
| 856 | 4 | 0 | |u http://dx.doi.org/10.7554/eLife.46314 |3 Volltext |
| 912 | |a GBV_USEFLAG_A | ||
| 912 | |a GBV_NLM | ||
| 951 | |a AR | ||
| 952 | |d 8 |j 2019 |b 24 |c 06 | ||