Controlling infectious pathogenic microbes, which pose a significant threat to human health, is increasingly crucial in various applications, including the medical sector, food packaging, and filtration membranes. In this study, a series of non-ionic antibacterial and antiviral copolymers incorporating eco-friendly cardanol moiety and short fluoroalkyl group are reported. The copolymers were synthesized via the radical copolymerization of methacrylate monomers, 2-hydroxyl-3-cardanylpropyl methacrylate and 1,3-bis(2,2,3,3,3-pentafluoropropoxy)propan-2-yl methacrylate. The incorporation of the fluoroalkyl group in the copolymer enhanced its hydrophobicity and reduced surface energy, indicating the improved stability in aqueous environments. Due to the crosslinkable point in cardanol, the copolymer series can be crosslinked by UV irradiation. The crosslinked polymer films demonstrated antibacterial (against Escherichia coli) and antiviral (against Emesvirus zinderi) properties. The fluoroalkyl moiety strengthens the antiviral properties, but in contrast, weakens the antibacterial properties. The physically stable and transparent crosslinked polymer films could be prepared as coatings on glass substrates or in free-standing form, and showed no cytotoxicity against human fibroblast cells. The polymer film in optimum content of the fluoroalkyl moiety (49 mol% of 1,3-bis(2,2,3,3,3-pentafluoropropoxy)propan-2-yl methacrylate unit in the copolymer) exhibited the outstanding antimicrobial properties, transparency, structural integrity after separated from the substrates, and low cytotoxicity.