title: Electrochemical detection of Nanog in cell extracts via target-induced resolution of an electrode-bound DNA pseudoknot.
abstract: Nanog is among the most important indicators of cell pluripotency and self-renew, so detection of Nanog is critical for tumor assessment and monitoring of clinical prognosis. In this work, a novel method for Nanog detection is proposed by using electrochemical technique based on target-induced conformational change of an electrode-bound DNA pseudoknot. In the absence of Nanog, the rigid structure of the pseudoknot will minimize the connection between the redox tag and the electrode, thus reducing the obtained faradaic current. Nevertheless, the Nanog binding may liberate the flexible single-stranded element that transforms the DNA pesudokont into DNA hairpin structure due to steric hindrance effect, thus making the electrochemical tag close to the electrode surface. Consequently, electron transfer can be enhanced and very well electrochemical response can be observed. By using the proposed method, Nanog can be determined in a linear range from 2nM to 25nM with a detection limit of 163 pM. Furthermore, the proposed method can be directly used to assay Nanog not only in purified samples but also in complex media (cell extracts), which shows potential applications in Nanog functional studies as well as clinical diagnosis in the future.
authors: State Key Laboratory of Pharmaceutical Biotechnology and Collaborative Innovation Center of Chemistry for Life Sciences, Department of Biochemistry, Nanjing University, Nanjing, 210093 PR China; State Key Laboratory of Reproductive Medicine, Department of Reproductive Health, Nanjing Maternity and Child Health Care Hospital affiliated with Nanjing Medical University, Nanjing, 210004 PR China.
authors: Jiehua
authors: J
authors: Ma
authors: State Key Laboratory of Pharmaceutical Biotechnology and Collaborative Innovation Center of Chemistry for Life Sciences, Department of Biochemistry, Nanjing University, Nanjing, 210093 PR China.
authors: Chao
authors: C
authors: Li
authors: State Key Laboratory of Pharmaceutical Biotechnology and Collaborative Innovation Center of Chemistry for Life Sciences, Department of Biochemistry, Nanjing University, Nanjing, 210093 PR China.
authors: Yaqin
authors: Y
authors: Tao
authors: State Key Laboratory of Pharmaceutical Biotechnology and Collaborative Innovation Center of Chemistry for Life Sciences, Department of Biochemistry, Nanjing University, Nanjing, 210093 PR China.
authors: Chang
authors: C
authors: Feng
authors: State Key Laboratory of Pharmaceutical Biotechnology and Collaborative Innovation Center of Chemistry for Life Sciences, Department of Biochemistry, Nanjing University, Nanjing, 210093 PR China; Center for Molecular Recognition and Biosensing, School of Life Sciences, Shanghai University, Shanghai, 200444 PR China. Electronic address: genxili@nju.edu.cn.
authors: Genxi
authors: G
authors: Li
mesh_terms: D000327:Adsorption; D003217:Conductometry / Q000295:instrumentation*; D015342:DNA Probes / Q000737:chemistry* / Q000648:ultrastructure; D004566:Electrodes; D004867:Equipment Design; D019544:Equipment Failure Analysis; D015336:Molecular Probe Techniques / Q000295:instrumentation*; D000071317:Nanog Homeobox Protein / Q000032:analysis* / Q000737:chemistry*; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity
chemical_list: D015342:DNA Probes; D000071317:Nanog Homeobox Protein
keywords: Cancer cells; Electrochemical biosensor; Nanog