Peripheral nerve injury (PNI) remains an unresolved challenge in the medicine area. With the development of biomaterial science and tissue engineering, a variety of nerve conduits were widely applied for repairing long defect PNI. DNA materials are developing rapidly due to their multiple advantages. In the present study, we aim to combine a DNA hydrogel, vascular endothelial growth factor (VEGF) and nerve growth factor (NGF) to construct a new type of delivery system, which could achieve a biphasic release profile of VEGF and NGF by taking advantage of the different degradation rates between X- and T-type DNA. In vitro results showed that the DNA gel + VEGF/NGF system could promote proliferation, migration and myelination of Rat Schwann cells (RSC) while maintaining cell viability. In vivo results indicated a better effect of DNA gel + VEGF/NGF on promoting repair of long defect PNI than the hollow chitin conduits (CT), DNA gel or VEGF/NGF group. The new technology invention holds promising clinical application prospects for repairing PNI and may be used broadly after step-by-step improvement.