Multifunctional Collagen‐Like Protein as a Gene Therapy Vehicle for Biomedical Applications

Gene therapy, utilizing a protein-based delivery system, offers a promising alternative to viral vectors owing to their stability, cell specificity, and adaptability. This study investigates the potential of collagen, a key extracellular matrix (ECM) component, as a gene delivery vehicle. While natural collagens play a crucial role in tissue regeneration, their high molecular weight limit efficient cellular uptake. To overcome this, we engineered a collagen-like protein (CLP) containing G-X-Y repeats mimicking type I collagen and retaining its functional properties. Furthermore, mussel adhesive protein (MAP) with histone-like characteristics is capable of delivering foreign genetic material into mammalian cells. We hypothesize that fusing CLP with MAP, would synergize the ECM-binding of collagen with the cationic property of MAP, enhancing gene delivery efficiency. The recombinant fusion protein was successfully expressed in E. coli and purified. The fusion protein exhibited strong DNA binding ability similar to MAP and the protein-DNA complex remained stable even in the presence of serum and DNase enzyme. The in vitro transfection studies substantiate the potential of CLP-MAP to deliver plasmid DNA in NIH-3T3 cells. Our findings indicate that the CLP-MAP fusion protein is effective in DNA binding and biocompatible, positioning it as a promising novel gene delivery system.