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SPECIFIC DETECTION OF RIBONUCLEIC ACID SEQUENCES USING NOVEL CRISPR ENZYME-MEDIATED DETECTION STRATEGIES

机译:使用新型CRISPR酶介导的检测策略对核糖核酸序列进行特异性检测

摘要

Embodiments disclosed herein include devices, methods, and systems for direct, selective, and sensitive detection of single-stranded target RNA sequences from various sources using a programmed Cas13a protein. When activated by binding a target RNA sequence, the Cas13a cleaves a tether releasing a reporter molecule that may then be detected. In some embodiments, the systems, methods, and devices may include a filter or membrane that may help to separate the tethered and untethered reporter molecules. These devices, systems, and techniques allow a user to rapidly process samples that may contain the target RNA, without needing to amplify the target sequences. These devices and methods may be used to assay a wide variety of samples and target RNA sources, for the presence or absence of a specific target RNA sequence. Compositions and kits, useful in practicing these methods, for example detecting a target RNA in a biological sample, are also described.
机译:本文公开的实施方案包括用于使用编程的Cas13a蛋白直接,选择性和灵敏地检测来自各种来源的单链靶RNA序列的装置,方法和系统。当通过结合靶RNA序列而被激活时,Cas13a会裂解一条链,释放出一个随后可被检测到的报道分子。在一些实施方案中,所述系统,方法和装置可以包括过滤器或膜,其可以帮助分离束缚的和未束缚的报道分子。这些设备,系统和技术使用户可以快速处理可能包含靶RNA的样品,而无需扩增靶序列。这些设备和方法可用于分析各种样品和靶RNA来源,以确定是否存在特定的靶RNA序列。还描述了可用于实施这些方法,例如检测生物样品中的靶RNA的组合物和试剂盒。

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