Using a transparent fracture replica with aperture size and water-cement ratio (w/c), the factors affecting the penetration behavior of rock grouting were investigated through laboratory experiments. In addition, the waterproof efficiency was estimated by the reduction of water outflow through the fractures after the grout curing process. Penetration behavior shows that grout penetration patterns present similarly radial forms in all experimental cases; however, velocity of grout penetration showed clear differences according to the aperture sizes and watercement ratio. It can be seen that the waterproof efficiency increased as the aperture size and w/c decreased. During grout injection or curing processes, air bubbles formed and bleeding occurred, both of which affected the waterproof ability of the grouting. These two phenomena can significantly prevent the successful performance of rock grouting in field-scale underground spaces, especially at deep depth conditions. Our research can provide a foundation for improving and optimizing the innovative techniques of rock grouting.