Orthogonal RNA imaging and simple RNA detection tools

国际上对RNA的调节作用的兴趣正在迅速增长。 以前被认为是“垃圾”DNA的基因组区域在基因调控中起着复杂的作用，通常被转录成RNA。 应激颗粒和其他动态细胞颗粒含有RNA，并被发现在基因调控中发挥重要作用。 然而，RNA研究工具仍处于萌芽状态，到2020年，RNA研究工具（不包括RNA干扰）的全球市场预计将达到近10亿美元（BCC研究）。 * 虽然蛋白质的研究通过多色荧光蛋白标签和基于正交蛋白质的亲和标签的发展而发生了革命性的变化，但RNA成像和纯化方法没有跟上步伐。 第一代RNA Mango在RNA成像和RNA蛋白复合物纯化方面具有很大的潜力，但目前还不允许多通道成像或正交纯化。 本NSERC CRD提案旨在直接解决这些问题，以便与应用生物材料（卑诗省伯纳比）一起继续快速商业化这项加拿大技术。该提案有两个具体目标：第一个是开发一个能够同时成像两个RNA的正交RNA Mango系统。 这些新的高亲和力适体将允许开发双色成像和正交纯化方法，并将具有许多体外和体内应用。 该提案的第二个目的是将第一代Mango技术应用于使用等温方法检测低浓度RNA。 总之，该提案将产生广泛的商业化元素，这将大大扩展第一代RNA Mango技术的实用性。

International interest in the regulatory role of RNA is rapidly growing. Genomic regions previously thought of as 'junk' DNA play complex roles in gene regulation and are typically transcribed into RNA. Stress granules and other dynamic cellular particles contain RNA and are being found to play important roles in gene regulation. Tools for the study of RNA however are still embryonic and the global market for RNA research tools (excluding RNA interference) is anticipated to reach nearly 1 billion US dollars by 2020 (BCC research). ****While the study of proteins was revolutionized by the development of multicolour fluorescent protein tags and orthogonal protein based affinity tags, RNA imaging and purification methods have not kept pace. The first generation of RNA Mango, holds much potential for RNA imaging and RNA protein complex purification, but currently does not allow multichannel imaging or orthogonal purifications. This NSERC CRD proposal aims to directly address these issues so as to continue the rapid commercialization of this Canadian technology together with Applied Biological Materials (Burnaby, BC).****The proposal has two specific aims: The first is to develop an orthogonal RNA Mango system capable of imaging two RNAs simultaneously. These new high affinity aptamers will allow the development of two colour imaging and orthogonal purification methodologies and will have many in vitro and in vivo applications. The second aim of this proposal will apply first generation Mango technology to the detect of low concentrations of RNA using isothermal methods. Together this proposal will generate a broad range of commercializable elements that will significantly extend the utility of first generation RNA Mango technology.