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When assessing student learning, it is pivotal that questions elicit ideal responses. This study focuses on the 9th grade STEMGenetics assessment item identification (e.g. MC, OR), creation, and rubric development based on the Knowledge Integration (KI) framework. Students participated in think-alouds and beta testing to determine the reasoning and cognitive demands required for the validity of assessment items, rubric development, and question refinement. This study focuses on questions that specifically align with our learning goals. This work provides transparency of the rigorous process of developing assessment items and rubrics that both elicit student responses and illuminates implications for school-settings that serve diverse populations.