Fabrication of stable and reversible DNA–RNA hammerhead ribozyme on a solid surface

Abstract

Although hammerhead ribozyme (HHRz) display various applications in solution, few studies have been developed based on surface immobilized HHRz mainly because RNA is highly degradable on surfaces. To overcome such limitations, here we report a stable HHRz fabricated on the surface of gold electrodes by a novel split DNA–RNA strategy, in which intact HHRz is split into two RNA portions and one of them is then replaced by a DNA segment. Mg²⁺-induced cleavage activity of the surface-tethered DNA–RNA HHRz was evaluated using electrochemical methods. It is found that the stability of the HHRz is remarkably improved after such replacement. Moreover, the structure and activity of the DNA–RNA HHRz can be regenerated very easily. In view of all these advantages, the DNA–RNA strategy is a promising alternative for the fabrication of ribozymes on surfaces, which also takes a firm step in advancing the applications of ribozymes on surfaces.