Integrating CRISPR-Cas9 Technology into Undergraduate Courses: Perspectives from a National Science Foundation (NSF) Workshop for Undergraduate Faculty, June 2018
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Authors:
Michael J. Wolyniak1,*,
Shane Austin2,
Lucian F. Bloodworth III1,
Dawn Carter3,
Scott H. Harrison4,
Tiffany Hoage5,
Lisa Hollis-Brown6,
Felicia Jefferson7,
Alison Krufka8,
Farida Safadi-Chamberlin9,
Maria S. Santisteban10,
Paula Soneral11,
Beth VanWinkle3,
Anil K. Challa12
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Received 22 October 2018 Accepted 23 February 2019 Published 26 April 2019
- ©2019 Author(s). Published by the American Society for Microbiology
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[open-access] This is an Open Access article distributed under the terms of the Creative Commons Attribution-Noncommercial-NoDerivatives 4.0 International license (https://creativecommons.org/licenses/by-nc-nd/4.0/ and https://creativecommons.org/licenses/by-nc-nd/4.0/legalcode), which grants the public the nonexclusive right to copy, distribute, or display the published work.
- *Corresponding author. Mailing address: Box 183, Hampden-Sydney, VA 23943. Phone: 434-223-6175. E-mail: [email protected].
Abstract:
As CRISPR (clustered regularly interspaced short palindromic repeats)-Cas9 technology becomes more mainstream in life science research, it becomes critical for undergraduate instructors to devise engaging ways to bring the technology into their classrooms. To help meet this challenge, the National Science Foundation sponsored a workshop for undergraduate instructors in June 2018 at The Ohio State University in conjunction with the annual Association of Biology Laboratory Educators meeting based on a workflow developed by the workshop’s facilitators. Over the course of two and a half days, participants worked through a modular workflow for the use of CRISPR-Cas9 in a course-based (undergraduate) research experience (CURE) setting while discussing the barriers each of their institutions had to implementing such work, and how such barriers could be overcome. The result of the workshop was a team with newfound energy and confidence to implement CRISPR-Cas9 technology in their courses and the development of a community of undergraduate educators dedicated to supporting each other in the implementation of the workflow either in a CURE or modular format. In this article, we review the activities and discussions from the workshop that helped each participant devise their own tailored approaches of how best to bring this exciting new technology into their classes.
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Abstract:
As CRISPR (clustered regularly interspaced short palindromic repeats)-Cas9 technology becomes more mainstream in life science research, it becomes critical for undergraduate instructors to devise engaging ways to bring the technology into their classrooms. To help meet this challenge, the National Science Foundation sponsored a workshop for undergraduate instructors in June 2018 at The Ohio State University in conjunction with the annual Association of Biology Laboratory Educators meeting based on a workflow developed by the workshop’s facilitators. Over the course of two and a half days, participants worked through a modular workflow for the use of CRISPR-Cas9 in a course-based (undergraduate) research experience (CURE) setting while discussing the barriers each of their institutions had to implementing such work, and how such barriers could be overcome. The result of the workshop was a team with newfound energy and confidence to implement CRISPR-Cas9 technology in their courses and the development of a community of undergraduate educators dedicated to supporting each other in the implementation of the workflow either in a CURE or modular format. In this article, we review the activities and discussions from the workshop that helped each participant devise their own tailored approaches of how best to bring this exciting new technology into their classes.

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Author and Article Information
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Received 22 October 2018 Accepted 23 February 2019 Published 26 April 2019
- ©2019 Author(s). Published by the American Society for Microbiology
-
[open-access] This is an Open Access article distributed under the terms of the Creative Commons Attribution-Noncommercial-NoDerivatives 4.0 International license (https://creativecommons.org/licenses/by-nc-nd/4.0/ and https://creativecommons.org/licenses/by-nc-nd/4.0/legalcode), which grants the public the nonexclusive right to copy, distribute, or display the published work.
- *Corresponding author. Mailing address: Box 183, Hampden-Sydney, VA 23943. Phone: 434-223-6175. E-mail: [email protected].
Figures

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FIGURE 1
Overview of the CRISPR-Cas9 lab workflow and correspondence with the flow of genetic information (Central Dogma of Molecular Biology). The yellow box shows the flow of genetic information culminating in the effect of gene function. The cream-colored boxes indicate bioinformatics. Boxes in dark blue indicate laboratory exercises, and boxes in light blue indicate variations and potential extensions for those lab exercises. CRISPR = clustered regularly interspaced short palindromic repeats; sgRNA = single guide RNA.

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FIGURE 2
Steps of the CRISPR-Cas9 workshop workflow. Phase 1 of the workflow involves gene analysis using a genome browser (e.g., ENSEMBL) and CRISPR/sgRNA design using bioinformatics tools (e.g., Benchling, CRISPRScan, ChopChop). In Phase 2, a double-stranded DNA (dsDNA) that will be used as a template for sgRNA synthesis is prepared. Phase 3 involves the synthesis of sgRNA by T7 RNA polymerase-driven in vitro transcription followed by quantitative and qualitative analysis of the synthesized RNA. During Phase 4, the nuclease activity of the sgRNA-Cas9 ribonucleoprotein complex on the PCR-amplified genomic target region is tested in vitro. CRISPR = clustered regularly interspaced short palindromic repeats; sgRNA = single guide RNA.

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FIGURE 3
Sample gels prepared and analyzed by undergraduate students in a summer course demonstrating in vitro nuclease assay of CRISPR-Cas9. The gel image in the left panel shows the efficacy of various sgRNA guides (C1–C4), along with various experimental controls to demonstrate the conditions required for optimal nuclease activity. The gel in the right panel shows nuclease activity at different concentrations of the sgRNA guide. CRISPR = clustered regularly interspaced short palindromic repeats; sgRNA = single guide RNA.

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FIGURE 4
Example of implementation plan in an undergraduate laboratory. sgRNA = single guide RNA; RT-PCR = reverse transcriptase PCR.