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Summary:* Bacillus subtilis spores have been identified as a promising platform for gene delivery and immunotherapy due to their exceptional resistance to environmental stresses, biocompatibility, and ease of manipulation.* Recent studies have demonstrated the potential of B. subtilis spore-based gene therapy in treating various types of cancer, including colon cancer, breast cancer, and pancreatic cancer.* B. subtilis spores can be loaded with genetic material using different methods such as electroporation, lipofection, and viral vectors, each having its advantages and limitations.* Researchers are exploring the potential of B. subtilis spores to deliver other therapeutic agents, such as siRNA and CRISPR/Cas9 systems, which could further enhance their immunotherapeutic potential.* While B. subtilis spore-based gene therapy has shown promising results, there are still challenges that need to be addressed, including the efficacy of gene delivery methods and the long-term stability and safety of B. subtilis spores as a delivery vehicle.* The potential applications of B. subtilis spores extend beyond cancer treatment to other diseases such as infectious diseases and genetic disorders.* Further research is needed to fully explore the potential of B. subtilis spores as a versatile platform for gene therapy and immunotherapy.