This article aims to develop a teachers’ perception scale regarding the artificial intelligence use in education. The scale development study was carried out in two stages during the 2023–2024 academic year, covering 597 teachers who stated that they used different artificial intelligence applications. Literature was thoroughly reviewed and focus group interviews were held with teachers who used artificial intelligence applications in education while pooling scale items. Field expert faculty members were consulted in evaluating face and content validity of the scale. Exploratory factor analysis was performed on the data obtained from the first sample group (n = 424), and a three-factor structure was determined in the first stage. It was observed that the factors of the first draft scale, consisting of 18 items, revealed 57.8% of the total variance. The first confirmatory factor analysis was conducted on the data collected from the second sample group (n = 173) in the second stage. It was confirmed that the structure consisting of 18 items and three factors (teaching perception, learning perception, and ethical perception) was compatible with the data. After the first-level confirmatory factor analysis for the Teachers’ Perception Scale Regarding Artificial Intelligence Use in Education, a second-level confirmatory factor analysis was conducted to determine whether the factors that made up the scale revealed the variable. The final scale, consisting of 15 items and three dimensions, was determined to be compatible with the data obtained. Reliability analysis presented that the Cronbach alpha internal consistency coefficient was calculated as .87 for the whole scale, .82 for learning perception, .79 for teaching perception, and .79 for ethical perception. The results show that the teachers’ perception scale regarding artificial intelligence use in education is valid and reliable, and a sound measurement tool to determine the perception regarding artificial intelligence use in education.