This study examined the perceptions of university faculty on their knowledge and accessibility practices in designing and delivering online courses for students with disabilities. An online survey developed by the researcher was utilized to collect data on perceptions of awareness of disability laws, quality standards, utilization of tools and professional development support. Results from the study indicate that professional development training in online learning had a very significant influence on accessibility knowledge and practice. In addition, high perceptions of knowledge for institutional policy and terminology and low perception for accessibility laws and standards were found. Given the results of the survey, this research study has implications for the type of training and professional development support faculty who teach online need to receive. It is also found that higher education institutions need to take a proactive stance regarding strengthening faculty knowledge and awareness of disability-related laws and regulations. This is to ensure that courses are developed and implemented to allow all students, including students with disabilities, to fully participate and engage online.