Abstract Peripheral nerve injury (PNI) is one of the common clinical injuries which needs to be addressed. Previous studies demonstrated the effectiveness of using biodegradable chitin (CT) conduits small gap tubulization technology as a substitute for traditional epineurial neurorrhaphy. Aiming to improve the effectiveness of CT conduits in repairing PNI, we modified their surface with a DNA-peptide coating. The coating consisted of single strand DNA (ssDNA) and its complementary DNAʹ-peptide mimics. First, we immobilize ssDNA (DNA1 + 2) on CT conduits by carbodiimide hydrochloride/N-hydroxysuccinimide (EDC/NHS) method to construct CT/DNA conduits. EDC/NHS was used to activate carboxyl groups of modified ssDNA for direct reaction with primary amines on the CT via amide bond formation. Then, DNA1ʹ-BDNF + DNA2ʹ-VEGF mimic peptide (RGI + KLT) were bonded to CT/DNA conduits by complementary base pairing principle at room temperature to form CT/RGI + KLT conduits. When the surrounding environment rose to a certain point (37 °C), the CT/RGI + KLT conduits achieved sustainable release of DNA′-peptide. In vitro, the CT conduits modified with the DNA-peptide coating promoted the proliferation and secretion of Schwann cells by maintaining their repair state. It also promoted the proliferation of human umbilical vein vessel endothelial cells and axon outgrowth of dorsal root ganglion explants. In vivo, CT/RGI + KLT conduits promoted regeneration of injured nerves and functional recovery of target muscles, which was facilitated by the synergistic contribution of angiogenesis and neurogenesis. Our research brings DNA and DNA-peptide hybrids into the realm of tissue engineering to repair PNI.