Rapid detection of methyltransferases utilizing dumbbell DNA-templated copper nanoparticles
Rapid detection of methyltransferases utilizing dumbbell DNA-templated copper nanoparticles
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DOI：
10.1016/j.snb.2018.08.140
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发表时间：
2018-12
期刊：
影响因子：
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通讯作者：
Jingxian Yin;Feng Liu;Tingting Fan;Yue Ren;Yuyang Jiang
中科院分区：
文献类型：
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作者：
Jingxian Yin;Feng Liu;Tingting Fan;Yue Ren;Yuyang Jiang
DNA甲基化作为DNA修饰的表观遗传事件，在遗传性疾病中发挥着重要作用，越来越多的证据表明DNA甲基转移酶（DNA methyltransferase，MTase）可作为遗传性疾病的生物标志物。基于目标保护原理，建立了一种DNA甲基转移酶的智能检测系统。在MTase存在下，哑铃状模板DNA（DT DNA）作为传感器被甲基化，并抵抗核酸内切酶和核酸外切酶的消化。因此，在上述检测系统中恢复Cu 2+后，能够快速检测荧光。然而，如果模板DNA在没有MTase保护的情况下被消化成核苷酸，则随后的荧光铜纳米颗粒（CuNP）合成步骤将失败，因为它是模板依赖性的。因此，在最佳条件下，可达到约1.6968 × 10−4U/μL MTase活性的检测限。此外，这种真实的样品分析显示出在复杂环境中的潜在应用前景。此外，所提出的MTase检测系统可用于筛选MTase的抑制剂或药物。
DNA methylation, as an epigenetic event of DNA modification, plays important roles in genetic disorders and diseases, such as cancer, and increasing evidence has shown that DNA methyltransferase (MTase) can be utilized as a biomarker for genetic diseases. In the present study, a smart detection system for DNA methyltransferase was established based on a target-protection principle. In the presence of MTase, as the sensor, dumbbell template DNA (DT DNA) was methylated and resisted endonuclease and exonuclease digestion. Thus, fluorescence was able to be rapidly detected after restoring Cu2+in the detection system above. However, if the template DNA was digested into nucleotides without the protection of MTase, the following step of fluorescence copper nanoparticle (CuNP) synthesis would fail because it is template dependent. Therefore, under optimal conditions, a detection limit of approximately 1.6968 × 10−4U/μL MTase activity was achieved. In addition, this real sample assay shows promise for potential application in complex environments. Furthermore, the proposed MTase-detection system can be utilized to screen inhibitors or drugs for MTase.