Vitamin B12 being a natural catalyst in atom transfer radical polymerization (ATRP), has the advantages of mild reaction conditions, good biocompatibility and high catalytic efficiency. In this report, an electrochemical biosensor of the lung cancer biomarker microRNA-21 (miRNA-21) is designed for early screening of lung cancer with high sensitivity at the femtomolar level. In this approach, hairpin DNA with N3 end group was first attached to the electrode surface. When miRNA-21 was present and paired with hairpin DNA, the N3 group released and attached to the ATRP initiator through "click reaction". Through eATRP, a large number of FerrocenylMethyl Methacrylate (FcMMA) monomers polymerized into long chains for signal amplification. These long chains had a distinct electrical signal in the square wave voltammetry (SWV), which can detect RNA with high sensitivity. The limit of detection (LOD) goes down to 1.010 fM after ATRP polymerization, which is lower than that of the majority of other ultra-sensitive RNA electrochemical assays. Results also show that the vitamin B12-based electrochemical biosensor is highly selective and suitable for RNA detection in complex biological samples.Copyright © 2023 Elsevier B.V. All rights reserved.