title: Carbon nanotube-DNA hybrid used for activity monitoring and inhibitor screening of nuclease.
abstract: Carbon nanotubes (CNTs) can efficiently quench the fluorescence of the adsorbed fluorophores and nonconvalently interact with soft single-stranded DNA (ssDNA). Upon disruption of CNTs-fluorescent oligonucleotides hybrid by nuclease S1, fluorescence turn-on was observed. Using this strategy, a platform based on fluorescence signal for monitoring the activity of nuclease with advantages of high sensitivity and commonality was established, and a linear relationship between initial cleavage reaction rate and nuclease S1 concentration is found in the range of 0.6-8.0 U mL(-1) with a detection limit of 0.08 U mL(-1). Furthermore, by taking pyrophosphate as an example, we use the assay to evaluate the prohibition effect on nuclease, and the extent of fluorescence recovery decreased linearly with increasing the concentration of pyrophosphate in the range of 0.2-1.4 mM, implying that the cleavage reaction by nuclease S1 was prohibited, and therefore this fluorescence assay can also be conveniently utilized for inhibitor screening of nuclease.
authors: Education Ministry Key Laboratory on Luminescence and Real-Time Analysis, College of Chemistry and Chemical Engineering, Southwest University, Chongqing, China.
authors: Zhong De
authors: ZD
authors: Liu
authors: Ping Ping
authors: PP
authors: Hu
authors: Heng Xin
authors: HX
authors: Zhao
authors: Yuan Fang
authors: YF
authors: Li
authors: Cheng Zhi
authors: CZ
authors: Huang
mesh_terms: D053837:DNA Cleavage; D004277:DNA, Single-Stranded / Q000737:chemistry / Q000378:metabolism*; D011756:Diphosphates / Q000737:chemistry; D004791:Enzyme Inhibitors / Q000737:chemistry*; D005456:Fluorescent Dyes / Q000737:chemistry; D007700:Kinetics; D037742:Nanotubes, Carbon / Q000737:chemistry*; D009841:Oligonucleotides / Q000138:chemical synthesis / Q000378:metabolism; D012235:Rhodamines / Q000737:chemistry; D015719:Single-Strand Specific DNA and RNA Endonucleases / Q000037:antagonists & inhibitors / Q000378:metabolism*; D013050:Spectrometry, Fluorescence
chemical_list: C114557:5-carboxytetramethylrhodamine succinimidyl ester; D004277:DNA, Single-Stranded; D011756:Diphosphates; D004791:Enzyme Inhibitors; D005456:Fluorescent Dyes; D037742:Nanotubes, Carbon; D009841:Oligonucleotides; D012235:Rhodamines; C107241:diphosphoric acid; D015719:Single-Strand Specific DNA and RNA Endonucleases