The present study aims to examine the effect of using Artificial Intelligence (AI) in translation training on assessment literacy and professional development among translation instructors. The study employed a quasi-experimental design using a pre-test/post-test control group. The statistical population consisted of all translation instructors in Shiraz, including faculty members, adjunct instructors, instructors at Islamic Azad University and other private/non-governmental universities, language-institute instructors who offered translation classes/workshops, instructors of translation test-preparation courses, and instructors teaching specialized translation. A convenience sample of 50 instructors was selected from this population and randomly assigned to an experimental group or a control group. Data were collected using a researcher-developed Assessment Literacy Questionnaire and Nova’s (2008) Professional Development Questionnaire. The data were analyzed using analysis of covariance (ANCOVA). The findings indicated that the use of AI tools in translation training affected the assessment literacy of translation instructors (p < 0.05). The use of AI tools in translation training also affected the professional development of translation instructors (p < 0.05). These results suggest that AI functions not only as a technological tool but also as a transformative factor in translation teaching–learning processes and in translation instructors’ professional growth. However, effective use of this capacity requires strengthening technological infrastructure, providing training in digital skills, and reinforcing ethical, data-informed policies in educational settings.