A Scalable CURE Using a CRISPR/Cas9 Fluorescent Protein Knock-In Strategy in Caenorhabditis elegans †

Genome editing with CRISPR/Cas9 technology has advanced from the lab bench to clinical application with multiple trials underway. This article introduces a course-based undergraduate experience (CURE) combining CRISPR/Cas9 genome editing (using a modified two-plasmid system) and the animal model Caenorhabditis elegans. This CURE is designed to be a scalable, semester-long laboratory that will introduce the students to literature searches, molecular biology, experiment planning, microscopy, CRISPR bioethics discussion, and scientific writing. Here, students challenged themselves to endogenously tag the C. elegans gene zmp-4, a matrix metalloproteinase enzyme, with a fluorescent protein marker and successfully generated a new worm strain. The knock-in was confirmed with genotyping and imaging and will be available for use by the entire worm community.