dc.contributor.author | Arbab, Mandana | |
dc.contributor.author | Mahony, Shaun | |
dc.contributor.author | Cho, Hyunjii | |
dc.contributor.author | Chick, Joel M. | |
dc.contributor.author | Rolfe, Philip Alexander | |
dc.contributor.author | Van Hoff, John Peter | |
dc.contributor.author | Morris, Viveca W. S. | |
dc.contributor.author | Gygi, Steven P. | |
dc.contributor.author | Maas, Richard L. | |
dc.contributor.author | Gifford, David K. | |
dc.contributor.author | Sherwood, Richard I. | |
dc.date.accessioned | 2013-03-22T21:06:45Z | |
dc.date.available | 2013-03-22T21:06:45Z | |
dc.date.issued | 2012-10 | |
dc.date.submitted | 2012-09 | |
dc.identifier.issn | 0305-1048 | |
dc.identifier.issn | 1362-4962 | |
dc.identifier.uri | http://hdl.handle.net/1721.1/77989 | |
dc.description.abstract | Interactions between DNA and transcription factors (TFs) guide cellular function and development, yet the complexities of gene regulation are still far from being understood. Such understanding is limited by a paucity of techniques with which to probe DNA–protein interactions. We have devised magnetic protein immobilization on enhancer DNA (MagPIE), a simple, rapid, multi-parametric assay using flow cytometric immunofluorescence to reveal interactions among TFs, chromatin structure and DNA. In MagPIE, synthesized DNA is bound to magnetic beads, which are then incubated with nuclear lysate, permitting sequence-specific binding by TFs, histones and methylation by native lysate factors that can be optionally inhibited with small molecules. Lysate protein–DNA binding is monitored by flow cytometric immunofluorescence, which allows for accurate comparative measurement of TF-DNA affinity. Combinatorial fluorescent staining allows simultaneous analysis of sequence-specific TF-DNA interaction and chromatin modification. MagPIE provides a simple and robust method to analyze complex epigenetic interactions in vitro. | en_US |
dc.language.iso | en_US | |
dc.publisher | Oxford University Press | en_US |
dc.relation.isversionof | http://dx.doi.org/10.1093/nar/gks1034 | en_US |
dc.rights | Creative Commons Attribution 3.0 | en_US |
dc.rights.uri | http://creativecommons.org/licenses/by-nc/3.0 | en_US |
dc.source | Oxford University Press | en_US |
dc.title | A multi-parametric flow cytometric assay to analyze DNA–protein interactions | en_US |
dc.type | Article | en_US |
dc.identifier.citation | Arbab, M. et al. “A Multi-parametric Flow Cytometric Assay to Analyze DNA-protein Interactions.” Nucleic Acids Research 41.2 (2012): e38–e38. | en_US |
dc.contributor.department | Massachusetts Institute of Technology. Computer Science and Artificial Intelligence Laboratory | en_US |
dc.contributor.department | Massachusetts Institute of Technology. School of Science | en_US |
dc.contributor.mitauthor | Mahony, Shaun | |
dc.contributor.mitauthor | Cho, Hyunjii | |
dc.contributor.mitauthor | Rolfe, Philip Alexander | |
dc.contributor.mitauthor | Gifford, David K. | |
dc.relation.journal | Nucleic Acids Research | en_US |
dc.eprint.version | Final published version | en_US |
dc.type.uri | http://purl.org/eprint/type/JournalArticle | en_US |
eprint.status | http://purl.org/eprint/status/PeerReviewed | en_US |
dspace.orderedauthors | Arbab, M.; Mahony, S.; Cho, H.; Chick, J. M.; Rolfe, P. A.; van Hoff, J. P.; Morris, V. W. S.; Gygi, S. P.; Maas, R. L.; Gifford, D. K.; Sherwood, R. I. | en |
dc.identifier.orcid | https://orcid.org/0000-0003-1709-4034 | |
mit.license | PUBLISHER_CC | en_US |
mit.metadata.status | Complete | |