Teacher knowledge of Autism Spectrum Disorder (ASD) plays a key role in the successful inclusion of children with the disorder in public schools. It therefore becomes important to understand what impacts upon teachers' knowledge of ASD. As such, we aimed to investigate the role of experience and culture on teachers' knowledge of ASD. Public teachers working in the United Kingdom (UK; n = 51) and People's Republic of China (n = 59) completed questionnaires measuring the experience of teaching children with ASD and levels of knowledge about the disorder. Analysis was then conducted to examine how knowledge of ASD differed as a function of culture and experience. Results showed that teachers who had experience of working with children with ASD demonstrated a higher level of knowledge relating to the disorder. Further, teachers in the UK had significantly more knowledge of ASD than teachers in China. The results suggest cross-cultural differences in teachers' understanding of ASD. Training opportunities which allow teachers to work with children with ASD should be provided in both the UK and in China.