Cultural differences between faculty and their students can create important challenges that affect the quality and efficacy of online teaching and learning. The objectives of this study were to: (a) create and pilot test an assessment for online faculty to measure culturally responsive teaching knowledge (CRT) and culturally responsive educational practices (CREP) in teaching and advising students of color, military students, LGBTQ students, religious minority students, and international students; (b) describe participants' CRT knowledge, value, and CREP; and (c) examine differences between their knowledge and their practices. The combined pilot and main study sample was comprised of 47 completed surveys. Internal consistency reliability was high for all subscales (.895-.970); subscale intra-class correlation coefficients ranged from .526-.833. The pilot round revealed strong face and content validity. Campbell-Fiske multitraitmultimethod matrix generated evidence of construct validity. Within-group comparisons of subscale scores using Wilcoxon Signed-ranks test revealed some significant differences between perceived knowledge and practice. Mann Whitney U test did not reveal significant differences in subscale scores or overall score by sector, degree level taught, or gender. The results have important implications for faculty training, professional development, mentoring, and support. Faculty who teach online who understand and value culturally responsive pedagogy and have the knowledge and skills to implement best practices in meeting the needs of diverse learners will enhance both teaching and learning. Culturally responsive knowledges and practice are particularly important as online programs are becoming ubiquitous across traditional institutions of higher education and their core faculty are teaching an increasingly diverse student body.