Sho Watanabe, Sangsin Lee, Manwal Harb, Shirin Nouraein, Emma Raisley, Honghao Li, Nicolas Buitrago, Beatrice Pforr, Jerzy O Szablowski
{"title":"Monitoring in vivo transcription with synthetic serum markers.","authors":"Sho Watanabe, Sangsin Lee, Manwal Harb, Shirin Nouraein, Emma Raisley, Honghao Li, Nicolas Buitrago, Beatrice Pforr, Jerzy O Szablowski","doi":"10.1101/2024.12.10.627810","DOIUrl":null,"url":null,"abstract":"<p><p>Understanding transcription profiles of living tissues is critical for biology and medicine. However, measurement of the transcript levels is typically done in homogenized tissues post-mortem. Here, we present a new platform that enables non-invasive monitoring of specific mRNA levels <i>in vivo</i> , without tissue destruction. We achieved this by combining two cutting-edge tools - synthetic serum markers, called Released Markers of Activity ( <b>RMAs</b> ), and RNA-based sensors of transcription. We call this platform IN-vivo Tracking of ACtive Transcription, or <b>INTACT</b> . In INTACT, when the target mRNA is expressed, the RNA sensor detects it and triggers the production and release of RMA reporters into the blood. Once in blood, the RMAs can be easily measured through a simple blood draw. Our data shows that INTACT can measure transcription of transgenes, as well as endogenous transcripts, such as <i>c-Fos</i> or <i>Arc</i> , both <i>in vivo</i> in the brain and in tissue culture. INTACT enables simple measurement of transcript level histories in genetically-targetable cell populations of living animals.</p>","PeriodicalId":519960,"journal":{"name":"bioRxiv : the preprint server for biology","volume":" ","pages":""},"PeriodicalIF":0.0000,"publicationDate":"2024-12-13","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11661152/pdf/","citationCount":"0","resultStr":null,"platform":"Semanticscholar","paperid":null,"PeriodicalName":"bioRxiv : the preprint server for biology","FirstCategoryId":"1085","ListUrlMain":"https://doi.org/10.1101/2024.12.10.627810","RegionNum":0,"RegionCategory":null,"ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"","JCRName":"","Score":null,"Total":0}
引用次数: 0
Abstract
Understanding transcription profiles of living tissues is critical for biology and medicine. However, measurement of the transcript levels is typically done in homogenized tissues post-mortem. Here, we present a new platform that enables non-invasive monitoring of specific mRNA levels in vivo , without tissue destruction. We achieved this by combining two cutting-edge tools - synthetic serum markers, called Released Markers of Activity ( RMAs ), and RNA-based sensors of transcription. We call this platform IN-vivo Tracking of ACtive Transcription, or INTACT . In INTACT, when the target mRNA is expressed, the RNA sensor detects it and triggers the production and release of RMA reporters into the blood. Once in blood, the RMAs can be easily measured through a simple blood draw. Our data shows that INTACT can measure transcription of transgenes, as well as endogenous transcripts, such as c-Fos or Arc , both in vivo in the brain and in tissue culture. INTACT enables simple measurement of transcript level histories in genetically-targetable cell populations of living animals.
了解活组织的转录谱对生物学和医学至关重要。然而,转录水平的测量通常是在死后均质组织中进行的。在这里,我们提出了一种新的平台,可以在不破坏组织的情况下对体内特定mRNA水平进行无创监测。我们通过结合两种尖端工具实现了这一目标——合成血清标记物,称为释放活性标记物(rma),以及基于rna的转录传感器。我们把这个平台称为IN-vivo Tracking of ACtive Transcription,简称为unchanged。在完好无损的情况下,当目标mRNA表达时,RNA传感器检测到它,并触发RMA报告蛋白的产生和释放到血液中。一旦进入血液,rma可以通过简单的抽血轻松测量。我们的数据表明,在大脑和组织培养中,完好无损可以测量转基因的转录,以及内源性转录物,如c-Fos或Arc。完好无损使转录水平的历史在活的动物的遗传靶向细胞群的简单测量。