Carbon-based　electrode　as　an　economically　benign　choice　is　widely　used　in　electrochemical　or　bio-electrochemical　systems.　However,　its　poor　electrical　conductivity　that　leading　high　overpotential　and　energy　loss,　especially　in　large　scale　facilities,　remains　a　bottleneck　for　its　application.　Herein,　a　unique　cross-stacking　multi-layered　reduced　graphene　oxide　(rGO)　and　polyaniline　(PANI)　modified　carbon　brush　is　fabricated　via　one-step　electro-depositing.　In　particular,　the　top-most　layer　of　the　modified　electrode　is　orientated　controlled　to　be　rGO　or　PANI　layer　by　adjusting　the　initial　CV　scanning　in　the　positive　or　negative　direction.　As　was　indicated　by　cyclic　voltammetry,　the　improved　electrochemical　activity　is　achieved　by　coupling　the　advantages　of　the　highly　conductive　network　offered　by　graphene　with　desirable　stability　provided　by　the　well-dispersed　deposition　of　nanoscale　PANI　particles.　In　comparison,　the　modified　electrode　with　rGO　on　the　top-most　layer　(LrGO)　showed　higher　degree　of　sp2-hybridized　-C-C-　ordered　structure　in　Raman　profile,　lower　O/C　ratio　in　XPS　analysis,　higher　Zeta　potential　(−2.05　mV)　and　more　hydrophilic　than　unmodified　one.　Moreover,　benefiting　from　the　unique　cross-stacking　multi-layered　matrix　of　rGO　and　PANI,　the　best　electrochemical　performance　was　achieved　on　the　electrode　LrGO　with　high　exerted　electrochemical　active　surface　area　(ECSA)　of　0.85　mF　cm−2,　and　the　charge　transfer　resistance　as　low　as　0.32　Ω.　The　findings　of　this　study　provide　a　guidance　for　the　modification　and　application　of　carbon-based　electrode　using　rGO　and　PANI,　which　potentially　enables　the　scaling-up　of　carbon-based　electrode　in　various　(bio-)　electrochemical　systems　with　high　electrochemical　performance.　©　2024　The　Authors