In this paper, the integration of directional antennas in a room-level received signal strength (RSS) fingerprinting-based indoor localization system (ILS) is studied. The sensor reader (SR), which is in charge of capturing the RSS to infer the tag position, can be attached to an omnidirectional or directional antenna. Unlike commonly-employed omnidirectional antennas, directional antennas can receive a stronger signal from the direction in which they are pointed, resulting in a different RSS distributions in space and, hence, more distinguishable fingerprints. A simulation tool and a system management software have been also developed to control the system and assist the initial antenna deployment, reducing time-consuming costs. A prototype was mounted in a real scenario, with a number of SRs with omnidirectional and directional antennas properly positioned. Different antenna configurations have been studied, evidencing a promising capability of directional antennas to enhance the performance of RSS fingerprinting-based ILS, reducing the number of required SRs and also increasing the localization success.