Bioadhesives or sealants have received considerable attention for suture-less wound sealing. However, most adhesive biomaterials involve complex molecular designs which lack dynamic mechanical properties, inherent hemostasis, as well as the capability of preventing postsurgical tissue adhesion. Herein, a nano-enabled supramolecular hydrogel sealant based on complementary DNA duplexes combined with multiple physicochemical crosslinks is designed and demonstrated. This supramolecular sealant exhibits excellent dynamic reversibility, low swelling, and wet adhesive properties. It can achieve rapid sealing of damaged tissues, blood coagulation, as well as hemostasis. Furthermore, DNA molecules endow the sealant with promising capabilities of preventing protein absorption, cell adhesion and postsurgical tissue adhesion. The in vivo evaluation based on the gastric perforation repair model shows that the sealant can improve granulation tissue growth, collagen deposition, and vascularization and facilitate gastric perforation closure. The nano-enabled DNA supramolecular hydrogel sealant represents a promising alternative for gastric perforation repair and has large clinical potential pertaining to suture-less repair of soft tissues. Data availabilityThe data reported in this manuscript are available upon request.