限制性酶
DNA
费斯特共振能量转移
核酸内切酶
酶
计算生物学
外源DNA
生物
转化(遗传学)
生物物理学
化学
生物化学
基因
物理
荧光
量子力学
作者
Susan Buckhout‐White,Chanel Person,Igor L. Medintz,Ellen R. Goldman
出处
期刊:ACS omega
[American Chemical Society]
日期:2018-01-17
卷期号:3 (1): 495-502
被引量:16
标识
DOI:10.1021/acsomega.7b01333
摘要
DNA nanostructures have been shown viable for the creation of complex logic-enabled sensing motifs. To date, most of these types of devices have been limited to the interaction with strictly DNA-type inputs. Restriction endonuclease represents a class of enzyme with endogenous specificity to DNA, and we hypothesize that these can be integrated with a DNA structure for use as inputs to trigger structural transformation and structural rearrangement. In this work, we reconfigured a three-arm DNA switch, which utilizes a cyclic Förster resonance energy transfer interaction between three dyes to produce complex output for the detection of three separate input regions to respond to restriction endonucleases, and investigated the efficacy of the enzyme targets. We demonstrate the ability to use three enzymes in one switch with no nonspecific interaction between cleavage sites. Further, we show that the enzymatic digestion can be harnessed to expose an active toehold into the DNA structure, allowing for single-pot addition of a small oligo in solution.
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