ANALYTICA CHIMICA ACTA
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2016
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IF 4.712
Pei Liu,Kexin Zhang,Ranran Zhang,Huanshun Yin,Yunlei Zhou,Shiyun Ai
Abnormal level of DNA methyltransferase (MTase) – mediated DNA methylation is closely related with cancer and bacterial diseases. Herein, a novel strategy based on the keypad lock of duplex DNA modified meso-SiO 2 @Fe 3 O 4 was developed for colorimetric assay of Dam MTase activity. When the Dam MTase was introduced, the duplex DNA can be methylated at a palindrome sequence of 5′-GATC-3′ and cleaved by the methylation-sensitive restriction endonuclease Dpn I. Due to the instability of the newly formed DNA fragment, the hybrid will separated into a single-stranded DNA. Then the keypad lock will open, and the catalytic reaction of TMB and H 2 O 2 can be initiated through the pores of meso-SiO 2 @Fe 3 O 4 , and a high color signal can be clearly observed by the naked eye. Contrarily, without Dam MTase, the catalytic reaction will not be initiated, and result no color signal. The proposed method exhibited a wide dynamic range with a low detection limit of 0.73 U/mL. Additionally, this way can be performed in human serum with satisfying recovery. And the inhibition of Dam MTase can also be well demonstrated by using paclitaxel as a model. Therefore, the designed way not only provides a platform for monitoring Dam MTase activity, but also useful for further application in disease diagnosis and drug discovery.
研究领域:
生物化学
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分子生物学
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癌症研究
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药物发现
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产品关联
DOI
PubMed
高影响因子
BIOSENSORS & BIOELECTRONICS
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2015
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IF 6.409
Pei Liu,Dandan Wang,Yunlei Zhou,Haiyan Wang,Huanshun Yin,Shiyun Ai
DNA methyltransferase (MTase) has received a large amount of attention due to its catalyzation of DNA methylation in both eukaryotes and prokaryotes , which has a close relationship to cancer and bacterial diseases. Herein, a novel electrochemical strategy based on Dpn I digestion triggering the combination of poly adenine (polyA) DNA with a gold nanoparticles functioned glassy carbon electrode (AuNPs/GCE), is developed for the simple and efficient detection of DNA MTase and inhibitor screening. Only one methylene blue (MB)-labeled DNA hairpin probe and two enzymes are involved in this designed method. In the presence of Dam MTase, the hairpin probe can be methylated and then cleaved by the restriction endonuclease . Thus, a MB-labeled polyA signal-stranded DNA product is introduced to the surface of AuNPs/GCE through the effect between polyA and AuNPs, resulting in an obvious electrochemical signal. On the contrary, in the absence of Dam MTase, the DNA probe cannot be cleaved and a relatively small electrochemical response can be observed. As a result, the as-proposed biosensor offered an efficient way for Dam MTase activity monitoring with a low detection of 0.27 U/mL, a wide linear range and good stability. Additionally, this assay holds great potential for further application in real biological matrices and inhibitors screening, which is expected to be useful in disease diagnosis and drug discovery.
研究领域:
生物化学
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分子生物学
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分析化学
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癌症研究
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微生物学
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产品关联
DOI
PubMed
高影响因子
ANALYTICAL CHEMISTRY
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2022
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IF 8.008
Fengzhe Li,Yingying Chen,Jinhua Shang,Qing Wang,Shizhen He,Xiwen Xing,Fuan Wang
Abnormal DNA methylation contributes to the annoying tumorigenesis and the elevated expression of methylation-related methyltransferase (MTase) is associated with many diseases. Hence DNA MTase could serve as a promising biomarker for cancer-specific diagnosis as well as a potential therapeutic target. Herein, we developed an isothermal autocatalytic hybridization reaction (AHR) circuit for the sensitive detection of MTase and its inhibitors by integrating the catalytic hairpin assembly (CHA) converter with the hybridization chain reaction (HCR) amplifier. The initiator-mediated HCR amplifier could generate amplified fluorescent readout, as well as numerous newly activated triggers for motivating the CHA converter. The CHA converter is designed to expose the identical sequence of HCR initiators that reversely powered the HCR amplifier. Thus, the trace amount of target could produce exponentially amplified fluorescent readout by the autocatalytic feedback cycle between HCR and CHA systems. Then an auxiliary hairpin was introduced to mediate the assay of Dam MTase via the well-established AHR circuit. The Dam MTase-catalyzed methylation of auxiliary hairpin leads to its subsequent efficient cleavage by DpnI endonuclease, thus resulting in the release of HCR initiators to initiate the AHR circuit. The programmable nature of the auxiliary hairpin allows its easy adaption into other MTase assay by simply changing the recognition site. This proposed AHR circuit permits a sensitive, robust, and versatile analysis of MTase with the limit of detection (LOD) of 0.011 U/mL. Lastly, the AHR circuit could be utilized for MTase analysis in real complex samples and for evaluating the cell-cycle-dependent expression of MTase. This developed MTase-sensing strategy holds promising potential for biomedical analysis and clinical diagnosis.
研究领域:
分子生物学
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生物医学工程
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癌症诊断
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生物化学
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产品关联
DOI
PubMed
高影响因子
BIOSENSORS & BIOELECTRONICS
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2018
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IF 8.173
Xiaolun Peng,Jinlin Zhu,Wei Wen,Ting Bao,Xiuhua Zhang,Hanping He,Shengfu Wang
In the paper, a triple-amplified biosensor was built for the purpose of ultrasensitive methyltransferase (Dam MTase) activity detection and inhibitor screening based on in-situ synthesized silver nanoclusters (AgNCs) as signal probes to provide amplified current signal coupling with gold nanoparticles/electrochemical reduced graphene oxide (AuNPs/ERGO) hybrids and hybridization chain reactions (HCR) amplification strategy. For biosensor preparation, the AuNPs/ERGO hybrids were firstly generated on the electrode surface by electrochemical co-reduction method, then the ds-DNA structures which comprised the specific recognition sequences (5′-G-A-T-C-3′) of the Dam MTase and the restrictive endonuclease DpnI were formed on the electrode by the hybridization of the assistant DNA with the capture DNA. The presence of Dam MTase methylated partial ds-DNA structures on the electrode which could be digested by DpnI and could not undergo HCR process. The unmethylated ds-DNA structures underwent HCR process to further hybridize with DNA1 and DNA2 to form ds-DNA superstructures as effective template for AgNCs in-situ synthesis. Oxidation peak current of silver was obtained as signal output for Dam MTase detection. Integrated synthesis and detection in one as well as combined metal nanocluster with DNA superstructures, this biosensor showed a good performance for Dam MTase activity detection with a detection limit of 0.0073 U/mL. Compared with reported biosensor, the designed biosensor showed high sensitivity and had potential in clinical diagnosis as well as inhibitor screening.
研究领域:
生物化学
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分析化学
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生物传感器
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分子生物学
✓ 已验证
产品关联
DOI
PubMed
高影响因子
危险品化学品经营许可证（带存储）