Super-Resolution Imaging of a 2.5 Kb Non-Repetitive DNA in situ in the Nuclear Genome Using Molecular Beacon Probes

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dc.contributor.ORCID0000-0002-7408-1830 (Zhang, MQ)en_US
dc.contributor.authorNi, Yanxiangen_US
dc.contributor.authorCao, Boen_US
dc.contributor.authorMa, Tszshanen_US
dc.contributor.authorNiu, Gangen_US
dc.contributor.authorHuo, Yingdongen_US
dc.contributor.authorHuang, Jiandongen_US
dc.contributor.authorChen, Dannien_US
dc.contributor.authorLiu, Yien_US
dc.contributor.authorYu, Binen_US
dc.contributor.authorZhang, Michael Q.en_US
dc.contributor.authorNiu, Hanbenen_US
dc.contributor.utdAuthorZhang, Michael Q.en_US
dc.date.accessioned2018-08-31T14:58:23Z
dc.date.available2018-08-31T14:58:23Z
dc.date.created2017-05-09en_US
dc.date.issued2018-08-31
dc.description.abstractHigh-resolution visualization of short non-repetitive DNA in situ in the nuclear genome is essential for studying looping interactions and chromatin organization in single cells. Recent advances in fluorescence in situ hybridization (FISH) using Oligopaint probes have enabled super resolution imaging of genomic domains with a resolution limit of 4.9 kb. To target shorter elements, we developed a simple FISH method that uses molecular beacon (MB) probes to facilitate the probe-target binding, while minimizing non-specific fluorescence. We used three-dimensional stochastic optical reconstruction microscopy (3D-STORM) with optimized imaging conditions to efficiently distinguish sparsely distributed Alexa-647 from background cellular autofluorescence. Utilizing 3D-STORM and only 29-34 individual MB probes, we observed 3D fine scale nanostructures of 2.5 kb integrated or endogenous unique DNA in situ in human or mouse genome, respectively. We demonstrated our MB-based FISH method was capable of visualizing the so far shortest non-repetitive genomic sequence in 3D at super-resolution.en_US
dc.description.departmentSchool of Natural Sciences and Mathematicsen_US
dc.description.sponsorshipNational Basic Research Program of China (2012CB825802), the National Natural Science Foundation of China (61235012, 31401146, 31361163004, 91019016,31671384), the Special-Funded Program on National Key Scientific Instru- ments and Equipment Development (2012YQ150092), the National Basic Research Program of China (2012CB316503, 2015CB352005), the National Natural Science Foundation of China (61178080, 61335001), and the Shenzhen Science and Technology Planning Project (JCYJ20150324141711698).en_US
dc.identifier.bibliographicCitationNi, Yanxiang, Bo Cao, Tszshan Ma, Gang Niu, et al. 2017. "Super-resolution imaging of a 2.5 kb non-repetitive DNA in situ in the nuclear genome using molecular beacon probes." eLife 6, doi:10.7554/eLife.21660en_US
dc.identifier.issn2050-084Xen_US
dc.identifier.urihttp://hdl.handle.net/10735.1/6037
dc.identifier.volume6en_US
dc.relation.urihttp://dx.doi.org/10.7554/eLife.21660
dc.rightsCC BY 4.0 (Attribution)en_US
dc.rights©2017 The Authorsen_US
dc.rights.urihttp://creativecommons.org/licenses/by/4.0/en_US
dc.sourceeLife
dc.subjectDNAen_US
dc.subjectFluorescence in situ hybridizationen_US
dc.subjectGenomesen_US
dc.subjectMicroscopyen_US
dc.subjectChromatinen_US
dc.titleSuper-Resolution Imaging of a 2.5 Kb Non-Repetitive DNA in situ in the Nuclear Genome Using Molecular Beacon Probesen_US
dc.type.genrearticleen_US

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