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dc.contributor.authorDuan, Yajian
dc.contributor.authorMa, Gaoen
dc.contributor.authorHuang, Xionggao
dc.contributor.authorD'Amore, Patricia Ann
dc.contributor.authorZhang, Feng
dc.contributor.authorLei, Hetian
dc.date.accessioned2018-02-23T20:52:30Z
dc.date.issued2016
dc.identifierQuick submit: 2017-06-23T14:42:40-0400
dc.identifier.citationDuan, Yajian, Gaoen Ma, Xionggao Huang, Patricia A. D’Amore, Feng Zhang, and Hetian Lei. 2016. “The Clustered, Regularly Interspaced, Short Palindromic Repeats-Associated Endonuclease 9 (CRISPR/Cas9)-createdMDM2T309G Mutation Enhances Vitreous-Induced Expression of MDM2 and Proliferation and Survival of Cells.” Journal of Biological Chemistry 291 (31) (May 31): 16339–16347. doi:10.1074/jbc.m116.729467.en_US
dc.identifier.issn0021-9258en_US
dc.identifier.urihttp://nrs.harvard.edu/urn-3:HUL.InstRepos:34865295
dc.description.abstractThe 309G allele of single nucleotide polymorphisms (SNPs) in the mouse double minute (MDM2) promoter locus is associated with a higher risk of cancer and proliferative vitreoretinopathy (PVR), but as to whether this SNP G309 contributes to the pathogenesis of PVR is to-date unknown. The clustered, regularly interspaced, short palindromic repeats (CRISPR)-associated endonuclease (Cas)9 from Streptococcus pyogenes (SpCas9) can be harnessed to manipulate a single or multiple nucleotides in mammalian cells. Here, we delivered SpCas9 and guide RNAs (SpGuides) using dual adeno-associated viral (AAV)-derived vectors to target the MDM2 genomic locus together with a homologous repair template for creating the mutation of MDM2 T309G in human primary retinal pigment epithelial (hPRPE) cells, whose genotype is MDM2 T309T. The next generation sequencing results indicated that there was 42.51% MDM2 G309 in the edited hPRPE cells using the AAV-CRISPR/Cas9. Our data showed that vitreous induced an increase in MDM2 and subsequent attenuation of p53 expression in the MDM2 T309G hPRPE cells. Furthermore, our experimental results demonstrated that the MDM2 T309G in the hPRPE cells enhanced vitreous-induced cell proliferation and survival, suggesting that this SNP contributes to the pathogenesis of PVR.en_US
dc.language.isoen_USen_US
dc.publisherAmerican Society for Biochemistry & Molecular Biology (ASBMB)en_US
dc.relation.isversionofdoi:10.1074/jbc.M116.729467en_US
dash.licenseLAA
dc.subjectCRISPRen_US
dc.subjectCas9en_US
dc.subjectMDM2 T309Gen_US
dc.subjectvitreousen_US
dc.subjectproliferationen_US
dc.subjectsurvivalen_US
dc.subjectapoptosisen_US
dc.subjectextracellular-signal-regulated kinase (ERK)en_US
dc.subjectepithelial cellen_US
dc.subjectgenetic polymorphismen_US
dc.subjectmolecular biologyen_US
dc.subjectcell growthen_US
dc.titleThe Clustered, Regularly Interspaced, Short Palindromic Repeats-associated Endonuclease 9 (CRISPR/Cas9)-created MDM2T309G Mutation Enhances Vitreous-induced Expression of MDM2 and Proliferation and Survival of Cellsen_US
dc.typeJournal Articleen_US
dc.date.updated2017-06-23T18:42:42Z
dc.description.versionVersion of Recorden_US
dc.relation.journalJournal of Biological Chemistryen_US
dash.depositing.authorD'Amore, Patricia Ann
dc.date.available2016
dc.date.available2018-02-23T20:52:30Z
dc.identifier.doi10.1074/jbc.M116.729467*
workflow.legacycommentscat.completeen_US
dash.authorsorderedfalse
dash.contributor.affiliatedLei, Hetian
dash.contributor.affiliatedD'Amore, Patricia


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