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dc.contributor.authorGao, Xin D.
dc.contributor.authorTu, Li-Chun
dc.contributor.authorMir, Aamir
dc.contributor.authorDing, Yue-He
dc.contributor.authorLeszyk, John D.
dc.contributor.authorDekker, Job
dc.contributor.authorShaffer, Scott A.
dc.contributor.authorZhu, Lihua Julie
dc.contributor.authorWolfe, Scot A.
dc.contributor.authorSontheimer, Erik J.
dc.date2022-08-11T08:08:23.000
dc.date.accessioned2022-08-23T15:53:17Z
dc.date.available2022-08-23T15:53:17Z
dc.date.issued2018-01-31
dc.date.submitted2018-06-14
dc.identifier.citation<p>bioRxiv 171819; doi: https://doi.org/10.1101/171819. <a href="https://doi.org/10.1101/171819" target="_blank">Link to preprint on bioRxiv service.</a></p>
dc.identifier.doi10.1101/171819
dc.identifier.urihttp://hdl.handle.net/20.500.14038/29323
dc.description.abstractMapping proteomic composition at distinct genomic loci and subnuclear landmarks in living cells has been a long-standing challenge. Here we report that dCas9-APEX2 Biotinylation at genomic Elements by Restricted Spatial Tagging (C-BERST) allows the rapid, unbiased mapping of proteomes near defined genomic loci, as demonstrated for telomeres and centromeres. By combining the spatially restricted enzymatic tagging enabled by APEX2 with programmable DNA targeting by dCas9, C-BERST has successfully identified nearly 50% of known telomere-associated factors and many known centromere-associated factors. We also identified and validated SLX4IP and RPA3 as telomeric factors, confirming C-BERST's utility as a discovery platform. C-BERST enables the rapid, high-throughput identification of proteins associated with specific sequences, facilitating annotation of these factors and their roles in nuclear and chromosome biology.
dc.language.isoen_US
dc.relation<p>Now published in <em>Nature Methods</em> doi: <a href="http://dx.doi.org/10.1038/s41592-018-0006-2" target="_blank">10.1038/s41592-018-0006-2</a>.</p>
dc.rightsThe copyright holder for this preprint (which was not peer-reviewed) is the author/funder. It is made available under a CC-BY 4.0 International license.
dc.rights.urihttp://creativecommons.org/licenses/by/4.0/
dc.subjectmolecular biology
dc.subjectdCas9-APEX2 Biotinylation at genomic Elements by Restricted Spatial Tagging
dc.subjectC-BERST
dc.subjectproteomes
dc.subjectgenomic loci
dc.subjectAmino Acids, Peptides, and Proteins
dc.subjectBiochemistry
dc.subjectGenetics and Genomics
dc.subjectMolecular Biology
dc.subjectStructural Biology
dc.titleC-BERST: Defining subnuclear proteomic landscapes at genomic elements with dCas9-APEX2 [preprint]
dc.typePreprint
dc.source.journaltitlebioRxiv
dc.identifier.legacyfulltexthttps://escholarship.umassmed.edu/cgi/viewcontent.cgi?article=2559&amp;context=faculty_pubs&amp;unstamped=1
dc.identifier.legacycoverpagehttps://escholarship.umassmed.edu/faculty_pubs/1549
dc.identifier.contextkey12317713
refterms.dateFOA2022-08-23T15:53:17Z
html.description.abstract<p>Mapping proteomic composition at distinct genomic loci and subnuclear landmarks in living cells has been a long-standing challenge. Here we report that d<strong>C</strong>as9-APEX2 <strong>B</strong>iotinylation at genomic <strong>E</strong>lements by <strong>R</strong>estricted <strong>S</strong>patial <strong>T</strong>agging (C-BERST) allows the rapid, unbiased mapping of proteomes near defined genomic loci, as demonstrated for telomeres and centromeres. By combining the spatially restricted enzymatic tagging enabled by APEX2 with programmable DNA targeting by dCas9, C-BERST has successfully identified nearly 50% of known telomere-associated factors and many known centromere-associated factors. We also identified and validated SLX4IP and RPA3 as telomeric factors, confirming C-BERST's utility as a discovery platform. C-BERST enables the rapid, high-throughput identification of proteins associated with specific sequences, facilitating annotation of these factors and their roles in nuclear and chromosome biology.</p>
dc.identifier.submissionpathfaculty_pubs/1549
dc.contributor.departmentDepartment of Molecular, Cell and Cancer Biology
dc.contributor.departmentDepartment of Biochemistry and Molecular Pharmacology
dc.contributor.departmentProgram in Systems Biology
dc.contributor.departmentProteomics and Mass Spectrometry Facility
dc.contributor.departmentRNA Therapeutics Institute


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The copyright holder for this preprint (which was not peer-reviewed) is the author/funder. It is made available under a CC-BY 4.0 International license.
Except where otherwise noted, this item's license is described as The copyright holder for this preprint (which was not peer-reviewed) is the author/funder. It is made available under a CC-BY 4.0 International license.