The invention discloses a one-component deoximation type room-temperature vulcanized silicone rubber sealant and a preparation method thereof. The sealant is prepared through the following steps: mixing alpha,omega-dyhydroxylpolydimethylsiloxane and a silane coupling agent at a high temperature, terminating alpha,omega-dyhydroxylpolydimethylsiloxane with the silane coupling agent, and then adding dimethyl silicone oil, a de-ketoxime type cross-linking agent, a filling material and a catalyst. Compared with a traditional deoximation type sealant, the sealant has excellent adhesive properties for stainless steel, aluminium alloy, magnesium alloy, PPO, PC, PVC, PMMA, ABS, PET and the like, and can be used for adhesion and sealing in the industries of electronics, automobiles, household appliances, photovoltaic modules and the like. Besides, the sealant has excellent water resistance and especially has a high strength retention rate under high-temperature and high-humidity conditions.