A hybrid gene carrier, HGP, has been successfully constructed through the genipin-mediated cross-linking of thermophilic histone and PEI25K. The thermophilic histone gene GK2215 was cloned from Geobacillus kastophilus HTA426 and overexpressed in Escherichia coli BL21. The thermophilic histone was systematically characterized and then cross-linked with PEI25K by genipin to obtain HGP. Notably, HGP exhibited superior transfection efficiency due to the synergistic effects between these two components: PEI25K mainly contributed to the condensation and transfer of pDNA, while thermophilic histone could enhance the endosomal escape and further nuclear location to achieve high gene expression. Meanwhile, HGP showed much lower cytotoxicity and hemolytic activity than PEI25K due to the introduction of nontoxic thermophilic histone. In addition, a strong intrinsic red fluorescence could be obviously observed in HGP. In conclusion, the protein-polymer hybrid carrier could potentially be used as a theranostic delivery system for achieving both efficient gene therapy and in vivo imaging.