Assess graduate-level understanding of how CRISPR-Cas9 recognizes DNA targets and the molecular, genomic, and experimental factors that determine off-target activity. Scope includes PAM recognition, guide RNA seed pairing and mismatch tolerance, R-loop formation and Cas9 conformational dynamics, binding/cleavage kinetics and thermodynamics, chromatin accessibility and DNA modifications, gRNA design principles, and methods for predicting, detecting, and mitigating off-targets; questions may ask for explanations, data interpretation, and evaluation of mitigation strategies.