Clostridium difficile (C. difficile) infection (CDI) is a significant health risk within hospital settings. C. difficile is an anaerobic gram-positive bacterium that spreads among people via the fecal-oral route. It forms spores and produces toxins in vivo, leading to hospital-acquired diarrhea and colitis, particularly in developed countries. The emergence and proliferation of C. difficile are likely due to the widespread use of antibiotics. This work proposed to develop a lipid nanoparticle-encapsulated, nucleoside-modified mRNA (mRNA-LNP) vaccine against CDI. The mRNA encoded one truncated fragment of the receptor binding domain (RBD) of C. difficile Toxin A. We assessed both humoral and cell-mediated immune responses in mice following immunization. Results showed that the vaccine induced strong humoral and cell-mediated immune responses. By conducting a spore challenge, we confirmed that the robust immune responses provided protection against CDI. This research highlights the potential for creating highly effective mRNA vaccines against bacterial pathogens and serves as a foundational model for future antibacterial mRNA vaccines.