Oncolytic virotherapy has become the frontier of biological therapy for tumor treatment in recent years, especially the Herpes Simplex Virus-1 (HSV-1) approved by the U.S. Food and Drug Administration (FDA). However, current commercial formulations using HSV-1 require strict storage conditions (-80°C) to cope with its unstable characteristics. In addition, direct injection of HSV-1 may lead to virus migration to nontumor tissues or cause unexpected immune responses representing a therapeutic risk. Herein, we developed an HSV-loaded lyotropic liquid crystal precursor (LLCP) system with in situ gelation properties as the vector for HSV-1. The prepared HSV-LLCP showed a rapid gelling property (within 2 s) and the shear viscosity ranged from 5 to 9 mPa·s, indicating its excellent drug delivery performance. Upon encapsulating in LLCP, the storage conditions were improved to -4°C within 28 days, and the HSV-1 was protected with its unique crystal lattice. The release behavior of HSV-1 LLCP showed a triphasic sustained-release pattern during the experiment period, avoiding the potential side effects. In addition, HSV-LLCP exhibited a superior oncolytic activity compared to HSV-1 solution in murine melanoma B16 cells. This study showed that HSV-LLCP would become an alternative and promising HSV-1 vector with high safety and stability for melanoma treatment in the clinic.