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CRISPR Gene Editing: Designing the gRNA and Donor Template

Author(s): Laurel Lorenz

Princeton

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Summary:
In this adaptation, students learn how CRISPR/Cas9 is used in bacterial immunity and gene editing. Students create both a gRNA target and a donor template to edit a gene. Mutations can be from the case study, Piwi Matter, or designed by the…

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In this adaptation, students learn how CRISPR/Cas9 is used in bacterial immunity and gene editing. Students create both a gRNA target and a donor template to edit a gene. Mutations can be from the case study, Piwi Matter, or designed by the instructor.

Description

In this adaptation of the case study, Piwi Matters, mid- to upper-level molecular biology students learn how bacteria use CRISPR/Cas9 as an immune system. In addition, students learn how scientists have adapted CRISPR/Cas9 to be used to make mutations in model organisms. By using published protocols, students design gRNA target sequences and a single-strand donor template to create the mutation that they designed in Piwi Matters. Alternatively instructors can have students design mutations of other genes. Finally, students present their intended mutation, gRNA target, and donor template in 2-4 minute oral presentations. Ultimately, the class uses these presentations to vote which mutation should be made.

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