Highly sensitive and reversible silicon nanowire biosensor to study nuclear hormone receptor protein and response element DNA interactions.
To thoroughly understand the role that estrogen receptors partake in regulation of gene expression, characterization of estrogen receptors (ERs) and estrogen-response elements (EREs) interactions is essential. In the work, we present a highly sensitive and reusable silicon nanowire (SiNW) biosensor to study the interactions between human ER proteins (ER, α and β subtypes) and EREs (dsDNA). The proteins were covalently immobilized on the SiNW surface. Various EREs including wild-type, mutant and scrambled DNA sequences were then applied to the protein-functionalized SiNW surface. Due to negatively charged dsDNA, binding of the EREs to the ERs on the n-type SiNW biosensor leads to the accumulation of negative charges on the surface, thereby inducing increase in resistance. The results show that the specificity of the ERE-ERα binding is higher than that of the ERE-ERβ binding, what is more, the mutant ERE reduces the binding affinity for both ERα and ERβ. By applying various concentrations of wild-type ERE to the bound ERα, a very low concentration of 10 fM wild-type ERE was found to be able to bind to the ERα. The reversible association and dissociation between ERα and wt-ERE was achieved, pointing to a reusable biosensor for protein-DNA binding. Through the study, we have established the SiNW biosensor as a promising method in providing comprehensive study for hormone receptor-response element interactions.
Zhang
Guo-Jun
GJ
Institute of Microelectronics, A*STAR (Agency for Science, Technology and Research), 11 Science Park Road, Singapore Science Park II, Singapore 117685, Singapore. zhanggj@ime.a-star.edu.sg
Huang
Min Joon
MJ
Luo
Zhan Hong Henry
ZH
Tay
Guang Kai Ignatius
GK
Lim
Eu-Jin Andy
EJ
Liu
Edison T
ET
Thomsen
Jane S
JS
D015374:Biosensing Techniques / Q000295:instrumentation*; D003217:Conductometry / Q000295:instrumentation*; D004247:DNA / Q000737:chemistry*; D004867:Equipment Design; D019544:Equipment Failure Analysis; D017743:Equipment Reuse; D036103:Nanotechnology / Q000295:instrumentation; D043942:Nanotubes / Q000737:chemistry* / Q000648:ultrastructure; D025941:Protein Interaction Mapping / Q000295:instrumentation*; D011960:Receptors, Estrogen / Q000737:chemistry*; D020218:Response Elements; D012680:Sensitivity and Specificity; D012825:Silicon / Q000737:chemistry*
D011960:Receptors, Estrogen; D004247:DNA; D012825:Silicon