Label-Free Sensitive Detection of DNA Methyltransferase by Target-Induced Hyperbranched Amplification with Zero Background Signal

Analytical Chemistry
Yan ZhangChun-yang Zhang

Abstract

DNA methyltransferases (MTases) may specifically recognize the short palindromic sequences and transfer a methyl group from S-adenosyl-l-methionine to target cytosine/adenine. The aberrant DNA methylation is linked to the abnormal DNA MTase activity, and some DNA MTases have become promising targets of anticancer/antimicrobial drugs. However, the reported DNA MTase assays often involve laborious operation, expensive instruments, and radio-labeled substrates. Here, we develop a simple and label-free fluorescent method to sensitively detect DNA adenine methyltransferase (Dam) on the basis of terminal deoxynucleotidyl transferase (TdT)-activated Endonuclease IV (Endo IV)-assisted hyperbranched amplification. We design a hairpin probe with a palindromic sequence in the stem as the substrate and a NH2-modified 3' end for the prevention of nonspecific amplification. The substrate may be methylated by Dam and subsequently cleaved by DpnI, producing three single-stranded DNAs, two of which with 3'-OH termini may be amplified by hyperbranched amplification to generate a distinct fluorescence signal. Because high exactitude of TdT enables the amplification only in the presence of free 3'-OH termini and Endo IV only hydrolyzes the intact ...Continue Reading

References

Mar 1, 1992·Journal of Bacteriology·E BoyeA Løbner-Olesen
Aug 11, 2001·Science·W ReikJ Walter
Sep 15, 2001·Nucleic Acids Research·X Cheng, R J Roberts
Feb 11, 2004·Analytical Biochemistry·Bo Yeon KimJong Seog Ahn
Apr 2, 2005·Current Opinion in Microbiology·Anders Løbner-OlesenMartin G Marinus
Feb 21, 2006·Nature Reviews. Microbiology·Didier Wion, Josep Casadesús
Feb 1, 2007·Analytical Chemistry·Jun LiXingwang Zhou
Feb 9, 2010·Nucleic Acids Research·Robert J WoodPeter L Roach
Sep 8, 2010·Cell Research·Vivek ShuklaChu-Xia Deng
Jan 14, 2011·Journal of Neuro-oncology·Ganeshkumar RajendranAnjali Shiras
Sep 6, 2012·Chemical Communications : Chem Comm·Tian TianXin Zhou
Feb 13, 2013·Nature Reviews. Genetics·Zachary D Smith, Alexander Meissner
Oct 30, 2013·Journal of the American Chemical Society·Natalie B Muren, Jacqueline K Barton
Jan 30, 2014·Analytical Chemistry·Huimin DengZhiqiang Gao
Jun 5, 2015·Chemical Communications : Chem Comm·Bao-Zhu ChiJian-Ding Qiu
Mar 10, 2017·Methods and Applications in Fluorescence·Yueying LiChun-Yang Zhang
Jun 13, 2017·Chemical Communications : Chem Comm·Fei MaChun-Yang Zhang

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Citations

Jun 14, 2019·Chemical Communications : Chem Comm·Yan ZhangChun-Yang Zhang
Jan 29, 2019·Artificial Cells, Nanomedicine, and Biotechnology·Bhagwan S BatuleHyun Gyu Park
Jan 7, 2019·Journal of Materials Chemistry. B, Materials for Biology and Medicine·Huige ZhangChun-Yang Zhang
May 1, 2018·Chemical Communications : Chem Comm·Chen-Chen LiChun-Yang Zhang
May 23, 2018·Luminescence : the Journal of Biological and Chemical Luminescence·Leiyi ZhangLunxi Duan

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