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dc.contributor.authorTomorsky, Johanna
dc.contributor.authorDeBlander, Leah
dc.contributor.authorKentros, Clifford
dc.contributor.authorDoe, Chris Q
dc.contributor.authorNiell, Cristopher M
dc.date.accessioned2018-07-30T06:57:59Z
dc.date.available2018-07-30T06:57:59Z
dc.date.created2018-01-04T21:13:55Z
dc.date.issued2017
dc.identifier.citationeNeuro. 2017, 4(5).nb_NO
dc.identifier.issn2373-2822
dc.identifier.urihttp://hdl.handle.net/11250/2506708
dc.description.abstractThiouracil (TU)-tagging is an intersectional method for covalently labeling newly transcribed RNAs within specific cell types. Cell type specificity is generated through targeted transgenic expression of the enzyme uracil phosphoribosyl transferase (UPRT); temporal specificity is generated through a pulse of the modified uracil analog 4TU. This technique has been applied in mouse using a Cre-dependent UPRT transgene, CA>GFPstop>HA-UPRT, to profile RNAs in endothelial cells, but it remained untested whether 4TU can cross the blood-brain barrier (BBB) or whether this transgene can be used to purify neuronal RNAs. Here, we crossed the CA>GFPstop>HA-UPRT transgenic mouse to a Sepw1-cre line to express UPRT in layer 2/3 of visual cortex or to an Nr5a1-cre line to express UPRT in layer 4 of visual cortex. We purified thiol-tagged mRNA from both genotypes at postnatal day (P)12, as well as from wild-type (WT) mice not expressing UPRT (background control). We found that a comparison of Sepw1-purified RNA to WT or Nr5a1-purified RNA allowed us to identify genes enriched in layer 2/3 of visual cortex. Here, we show that Cre-dependent UPRT expression can be used to purify cell type-specific mRNA from the intact mouse brain and provide the first evidence that 4TU can cross the BBB to label RNA in vivo.nb_NO
dc.language.isoengnb_NO
dc.publisherSociety for Neurosciencenb_NO
dc.rightsNavngivelse 4.0 Internasjonal*
dc.rights.urihttp://creativecommons.org/licenses/by/4.0/deed.no*
dc.titleTU-tagging: A method for identifying layer-enriched neuronal genes in developing mouse visual cortexnb_NO
dc.title.alternativeTU-tagging: A method for identifying layer-enriched neuronal genes in developing mouse visual cortexnb_NO
dc.typeJournal articlenb_NO
dc.typePeer reviewednb_NO
dc.description.versionpublishedVersionnb_NO
dc.source.volume4nb_NO
dc.source.journaleNeuronb_NO
dc.source.issue5nb_NO
dc.identifier.doi10.1523/ENEURO.0181-17.2017
dc.identifier.cristin1536246
dc.description.localcodeCopyright © 2017 Tomorsky et al. This is an open-access article distributed under the terms of the Creative Commons Attribution 4.0 International license, which permits unrestricted use, distribution and reproduction in any medium provided that the original work is properly attributed.nb_NO
cristin.unitcode194,65,60,0
cristin.unitnameKavliinstitutt for nevrovitenskap
cristin.ispublishedtrue
cristin.fulltextoriginal
cristin.qualitycode1


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