Room temperature (RT) synthesis of high-performance zeolite membranes, which is profound from techno-economic and eco-friendly perspectives, remains a grand challenge. In this work, we pioneered the RT preparation of well-intergrown pure-silica MFI zeolite (Si-MFI) membranes, which was realized through adopting highly reactive NH4F-mediated gel as nutrient during epitaxial growth. Benefiting from the introduction of fluoride anions as mineralizing agent as well as precisely tuned nucleation and growth kinetics at RT, both their grain boundary structure and thickness could be deliberately controlled, resulting in the formation of Si-MFI membranes showing unprecedented n-/i-butane separation factor (96.7) and n-butane permeance (5.16 × 10-7 mol m-2 s-1 Pa-1) in the case of a feed molar ratio of 10/90, which well transcended the state-of-the-art membranes reported in the literature. This RT synthetic protocol was also proven effective for preparing highly b-oriented Si-MFI film, thus showing great promise for the preparation of diverse zeolite membranes with optimized microstructure and superior performance.