In the vanguard of biomedical innovation, regenerative medicine emerges as a transformative paradigm, concentrating its efforts on rectifying tissue deficits and functional aberrations in patients through the strategic augmentation of endogenous cellular processes. Central to this approach is nucleic acid therapy, which offers a novel pathway for tissue regeneration by modulating key signaling pathways. As one of the most effective ways to regulate cell function, nucleic acids are crucial for various tissue regeneration; however, their in vivo therapeutic applications face substantial challenges, including nuclease degradation, cell membrane impermeability, and targeted intracellular transport. Biomaterial-based gene delivery systems offer a solution for stable and localized drug delivery by enabling the controlled overexpression of therapeutic nucleic acids, producing functional regulatory agents. This review presents examples of nucleic acid applications in regenerative medicine, highlighting the synergy between nucleic acids and biomaterial technologies. It underlines the importance of nucleic acid delivery techniques, the choice of therapeutic nucleic acids, and biomaterials for advancing tissue repair. The latest developments in designing nucleic acid biomaterial-based delivery vehicles are explored, the mechanism of nucleic acids in tissue regeneration is elucidated, and their limitations are discussed while considering future directions for the clinical translation of nucleic acid-based therapeutics.