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Telomerase Triggered DNA Walker with a Superhairpin Structure for Human Telomerase Activity Sensing.
Analytical Chemistry ( IF 6.7 ) Pub Date : 2019-05-17 00:00:00 , DOI: 10.1021/acs.analchem.9b01603 Jing Huang 1 , Longyi Zhu 1 , Huangxian Ju 1 , Jianping Lei 1
Analytical Chemistry ( IF 6.7 ) Pub Date : 2019-05-17 00:00:00 , DOI: 10.1021/acs.analchem.9b01603 Jing Huang 1 , Longyi Zhu 1 , Huangxian Ju 1 , Jianping Lei 1
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Different from conventional DNA walkers, we designed a telomerase-triggered three-dimensional DNA walker consisting of a superhairpin structure with a bulged loop in the stem as the walking strand and dye-labeled tracks for ultrasensitive detection of telomerase activity. In the presence of telomerase, the primers in the stem of the superhairpin structure were elongated and triggered internal strand displacement, thus activating the superhairpin structure. Subsequently, the open superhairpin structure as a swing arm was able to bind with the track, and the swing arm could be released by enzymatic cleavage of the binding duplex domain, resulting in the fluorescence recovery of dye-labeled fragments from the surface of gold nanoparticles. Based on signal amplification of the telomerase-triggered DNA walker, the walking device was further applied to various cancer lines with a low detection limit of telomerase activity equivalent to 90 cells μL–1 for HeLa cells. Moreover, the advantage of this DNA walker strategy was confirmed by calculating telomerase activity in a single cell. This telomerase-triggered DNA walker provides a new concept on signal transduction for telomerase detection and is anticipated to stimulate interest in DNA nanomachine design for bioanalysis.
中文翻译：
具有人发端粒酶活性传感的超级发夹结构的端粒酶触发的DNA Walker。
与传统的DNA Walker不同，我们设计了一种端粒酶触发的三维DNA Walker，该结构由一个超级发夹结构组成，茎中有一个凸起的环作为行走链，并带有染料标记的轨道，可用于超灵敏地检测端粒酶活性。在端粒酶存在下，超级发夹结构茎中的引物被拉长并触发内部链移位，从而激活超级发夹结构。随后，开放的超级发夹结构作为摆臂能够与轨道结合，并且可以通过酶切裂解结合的双链结构域来释放摆臂，从而从金纳米颗粒的表面荧光回收染料标记的片段。根据端粒酶触发的DNA Walker的信号放大，–1用于HeLa细胞。此外，通过计算单个细胞中的端粒酶活性，证实了这种DNA Walker策略的优势。这种端粒酶触发的DNA Walker为端粒酶检测提供了信号转导的新概念，并有望激发人们对DNA纳米机设计进行生物分析的兴趣。
更新日期：2019-05-17
中文翻译：
具有人发端粒酶活性传感的超级发夹结构的端粒酶触发的DNA Walker。
与传统的DNA Walker不同，我们设计了一种端粒酶触发的三维DNA Walker，该结构由一个超级发夹结构组成，茎中有一个凸起的环作为行走链，并带有染料标记的轨道，可用于超灵敏地检测端粒酶活性。在端粒酶存在下，超级发夹结构茎中的引物被拉长并触发内部链移位，从而激活超级发夹结构。随后，开放的超级发夹结构作为摆臂能够与轨道结合，并且可以通过酶切裂解结合的双链结构域来释放摆臂，从而从金纳米颗粒的表面荧光回收染料标记的片段。根据端粒酶触发的DNA Walker的信号放大，–1用于HeLa细胞。此外，通过计算单个细胞中的端粒酶活性，证实了这种DNA Walker策略的优势。这种端粒酶触发的DNA Walker为端粒酶检测提供了信号转导的新概念，并有望激发人们对DNA纳米机设计进行生物分析的兴趣。
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