Orthogonal RNA imaging and simple RNA detection tools

国际上对RNA调控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，在基因调控中起着重要作用。然而，RNA研究工具仍处于萌芽阶段，RNA研究工具（不包括RNA干扰）的全球市场预计到2020年将达到近10亿美元（BCC研究）。****随着多色荧光蛋白标签和基于正交蛋白的亲和标签的发展，蛋白质的研究发生了革命性的变化，但RNA成像和纯化方法却没有跟上。第一代RNA芒果具有RNA成像和RNA蛋白复合物纯化的潜力，但目前不允许多通道成像或正交纯化。这项NSERC CRD提案旨在直接解决这些问题，以便与应用生物材料公司（BC省本拿比）一起继续这项加拿大技术的快速商业化。****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芒果系统。这些新的高亲和力适配体将允许双色成像和正交纯化方法的发展，并将有许多体外和体内应用。本提案的第二个目标是将第一代Mango技术应用于使用等温方法检测低浓度RNA。该提案将产生广泛的可商业化元件，这将大大扩展第一代RNA芒果技术的实用性。

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.