Using CRISPR-cas9 as a tool to generate double stranded breaks in yeast, students can explore the processes involved in DNA repair. We describe an experimental package that allows students to learn valuable tool building skills but with an emphasis on hypothesis driven research and experimental design. This platform inspires students to generate diverse hypotheses and allows for the collection of unique data sets for analysis. Our approach is to target the ADE2 gene that has a distinct red colour for loss of function alleles. Students design and utilse homology directed repair (HDR) templates to create a wide range of edits to the ADE2 gene; this can range from point mutations to much larger insertions and deletions. Students take ownership of their designs, and gain confidence to plan, implement, and execute a real experiment.We will review several examples of experimental predictions and outcomes that have allowed students to see the scientific process unfold. Students make unanticipated discoveries that lead to fertile discussions regarding models of DNA repair and CRISPR function. Through data analysis, students refine their understanding and challenge their assumptions, and this is a launchpad for conversations to consider more applied applications for gene therapy.
Keywords: DNA repair, inquiry, experimental design.
University of Manitoba (2025)
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