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Single-cell mechanogenetics using monovalent magnetoplasmonic nanoparticles

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dc.contributor.authorJi-wook Kim-
dc.contributor.authorSeo, D-
dc.contributor.authorJung-uk Lee-
dc.contributor.authorSouthard, KM-
dc.contributor.authorYongjun Lim-
dc.contributor.authorDaehyun Kim-
dc.contributor.authorGartner, ZJ-
dc.contributor.authorYoung-wook Jun-
dc.contributor.authorJinwoo Cheon-
dc.date.available2018-01-09T07:12:17Z-
dc.date.created2017-09-25-
dc.date.issued2017-09-
dc.identifier.issn1754-2189-
dc.identifier.urihttps://pr.ibs.re.kr/handle/8788114/4200-
dc.description.abstractSpatiotemporal interrogation of signal transduction at the single-cell level is necessary to answer a host of important biological questions. This protocol describes a nanotechnology-based single-cell and single-molecule perturbation tool, termed mechanogenetics, that enables precise spatial and mechanical control over genetically encoded cell-surface receptors in live cells. The key components of this tool are a magnetoplasmonic nanoparticle (MPN) actuator that delivers defined spatial and mechanical cues to receptors through target-specific one-to-one engagement and a micromagnetic tweezers (mu MT) that remotely controls the magnitude of force exerted on a single MPN. In our approach, a SNAP-tagged cell-surface receptor of interest is conjugated with a single-stranded DNA oligonucleotide, which hybridizes to its complementary oligonucleotide on the MPN. This protocol consists of four major stages: (i) chemical synthesis of MPNs, (ii) conjugation with DNA and purification of monovalent MPNs, (iii) modular targeting of MPNs to cell-surface receptors, and (iv) control of spatial and mechanical properties of targeted mechanosensitive receptors in live cells by adjusting the mu MT-to-MPN distance. Using benzylguanine (BG)-functionalized MPNs and model cell lines expressing either SNAP-tagged Notch or vascular endothelial cadherin (VE-cadherin), we provide stepwise instructions for mechanogenetic control of receptor clustering and for mechanical receptor activation. The ability of this method to differentially control spatial and mechanical inputs to targeted receptors makes it particularly useful for interrogating the differential contributions of each individual cue to cell signaling. The entire procedure takes up to 1 week-
dc.description.uri1-
dc.language영어-
dc.publisherNATURE PUBLISHING GROUP-
dc.titleSingle-cell mechanogenetics using monovalent magnetoplasmonic nanoparticles-
dc.typeArticle-
dc.type.rimsART-
dc.identifier.wosid000408997500003-
dc.identifier.scopusid2-s2.0-85028501864-
dc.identifier.rimsid60178ko
dc.date.tcdate2018-10-01-
dc.contributor.affiliatedAuthorJi-wook Kim-
dc.contributor.affiliatedAuthorJung-uk Lee-
dc.contributor.affiliatedAuthorYongjun Lim-
dc.contributor.affiliatedAuthorDaehyun Kim-
dc.contributor.affiliatedAuthorYoung-wook Jun-
dc.contributor.affiliatedAuthorJinwoo Cheon-
dc.identifier.doi10.1038/nprot.2017.071-
dc.identifier.bibliographicCitationNATURE PROTOCOLS, v.12, no.9, pp.1871 - 1889-
dc.citation.titleNATURE PROTOCOLS-
dc.citation.volume12-
dc.citation.number9-
dc.citation.startPage1871-
dc.citation.endPage1889-
dc.date.scptcdate2018-10-01-
dc.description.wostc3-
dc.description.scptc3-
dc.embargo.liftdate9999-12-31-
dc.embargo.terms9999-12-31-
dc.description.journalClass1-
dc.description.journalRegisteredClassscie-
dc.description.journalRegisteredClassscopus-
dc.subject.keywordPlusMOLECULE FORCE SPECTROSCOPY-
dc.subject.keywordPlusMAGNETIC NANOPARTICLES-
dc.subject.keywordPlusLIVING CELLS-
dc.subject.keywordPlusINTRACELLULAR DELIVERY-
dc.subject.keywordPlusSPATIOTEMPORAL CONTROL-
dc.subject.keywordPlusSIGNAL-TRANSDUCTION-
dc.subject.keywordPlusENDOSOMAL ESCAPE-
dc.subject.keywordPlusFE NANOPARTICLES-
dc.subject.keywordPlusREMOTE-CONTROL-
dc.subject.keywordPlusQUANTUM DOTS-
Appears in Collections:
Center for Nanomedicine (나노의학 연구단) > 1. Journal Papers (저널논문)
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