利用 RNA 整合子检测活细胞中的低丰度代谢物。
Detection of Low-Abundance Metabolites in Live Cells Using an RNA Integrator.
机构信息
Department of Chemistry, University of Massachusetts, Amherst, MA 01003, USA; Department of Pharmacology, Weill Medical College, Weill Cornell Medicine, New York, NY 10065, USA.
Department of Pharmacology, Weill Medical College, Weill Cornell Medicine, New York, NY 10065, USA.
出版信息
Cell Chem Biol. 2019 Apr 18;26(4):471-481.e3. doi: 10.1016/j.chembiol.2019.01.005. Epub 2019 Feb 14.
Genetically encoded biosensors are useful tools for detecting the presence and levels of diverse biomolecules in living cells. However, low-abundance targets are difficult to detect because they are often unable to bind and activate enough biosensors to detect using standard microscopic imaging approaches. Here we describe a type of RNA-based biosensor, an RNA integrator, which enables detection of low-abundance targets in vitro and in living cells. The RNA integrator is an RNA sequence comprising a ribozyme and an unfolded form of the fluorogenic aptamer Broccoli. Upon binding its target, the ribozyme undergoes cleavage and releases Broccoli, which subsequently folds and becomes fluorescent. Importantly, each target molecule can bind and induce cleavage of multiple copies of the integrator sensor, resulting in an amplified signal. We show that this approach can be generalized to numerous different ribozyme types for the detection of various small molecules.
基因编码生物传感器是检测活细胞中各种生物分子存在和水平的有用工具。然而，由于低丰度靶标通常无法结合并激活足够数量的生物传感器，因此难以检测。在这里，我们描述了一种基于 RNA 的生物传感器，即 RNA 整合子，它可以在体外和活细胞中检测低丰度靶标。RNA 整合子是一种由核酶和荧光适体 Broccoli 的未折叠形式组成的 RNA 序列。与靶标结合后，核酶会发生切割并释放 Broccoli，Broccoli 随后折叠并变得荧光。重要的是，每个靶分子都可以结合并诱导多个整合子传感器的切割，从而产生放大的信号。我们表明，这种方法可以推广到许多不同的核酶类型，用于检测各种小分子。
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