Configuring robust DNA strand displacement reactions forﾠin situﾠmolecular analyses

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dc.citation.firstpage3289
dc.citation.issueNumber7
dc.citation.journalTitleNucleic Acids Research
dc.citation.lastpage3298
dc.citation.volumeNumber40
dc.contributor.authorDuose, Dzifa Y.
dc.contributor.authorSchweller, Ryan M.
dc.contributor.authorZimak, Jan
dc.contributor.authorRogers, Arthur R.
dc.contributor.authorHittelman, Walter N.
dc.contributor.authorDiehl, Michael R.
dc.date.accessioned2016-01-29T18:44:40Z
dc.date.available2016-01-29T18:44:40Z
dc.date.issued2012
dc.description.abstractThe number of distinct biomolecules that can be visualized within individual cells and tissue sections via fluorescence microscopy is limited by the spectral overlap of the fluorescent dye molecules that are coupled permanently to their targets. This issue prohibits characterization of important functional relationships between different molecular pathway components in cells. Yet, recent improved understandings of DNA strand displacement reactions now provides opportunities to create programmable labeling and detection approaches that operate through controlled transient interactions between different dynamic DNA complexes. We examined whether erasable molecular imaging probes could be created that harness this mechanism to couple and then remove fluorophore-bearing oligonucleotides to and from DNA-tagged protein markers within fixed cell samples. We show that the efficiency of marker erasing via strand displacement can be limited by non-toehold mediated stand exchange processes that lower the rates that fluorophore-bearing strands diffuse out of cells. Two probe constructions are described that avoid this problem and allow efficient fluorophore removal from their targets. With these modifications, we show one can at least double the number of proteins that can be visualized on the same cells via reiterativein situﾠlabeling and erasing of markers on cells.
dc.identifier.citationDuose, Dzifa Y., Schweller, Ryan M., Zimak, Jan, et al.. "Configuring robust DNA strand displacement reactions forﾠin situﾠmolecular analyses." <i>Nucleic Acids Research,</i> 40, no. 7 (2012) Oxford University Press: 3289-3298. http://dx.doi.org/10.1093/nar/gkr1209.
dc.identifier.doihttp://dx.doi.org/10.1093/nar/gkr1209
dc.identifier.urihttps://hdl.handle.net/1911/88268
dc.language.isoeng
dc.publisherOxford University Press
dc.rightsThis is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License, which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.
dc.rights.urihttps://creativecommons.org/licenses/by-nc/3.0/us/
dc.titleConfiguring robust DNA strand displacement reactions forﾠin situﾠmolecular analyses
dc.typeJournal article
dc.type.dcmiText
dc.type.publicationpublisher version
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