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Integrating CRISPR-Cas9 Technology into Undergraduate Courses: Perspectives from a National Science Foundation (NSF) Workshop for Undergraduate Faculty, June 2018

    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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    Affiliations: 1: Department of Biology, Hampden-Sydney College, Hampden-Sydney, VA 23943; 2: Department of Biological and Chemical Sciences, The University of the West Indies at Cave Hill, Bridgetown, BB11000, Barbados; 3: Thomas H. Gosnell School of Life Sciences, Rochester Institute of Technology, Rochester, NY 14623; 4: Department of Biology, North Carolina A&T State University, Greensboro, NC 27411; 5: Department of Biology, University of Wisconsin—Stout, Menomonie, WI 54751; 6: Department of Biology, Pikes Peak Community College, Colorado Springs, CO 80906; 7: Department of Biology, Fort Valley State University, Fort Valley, GA 31030; 8: Department of Biological Sciences, Rowan University, Glassboro, NJ 08028; 9: Department of Biochemistry and Molecular Biology, Colorado State University, Fort Collins, CO 80523; 10: Department of Biology, University of North Carolina at Pembroke, Pembroke, NC 28372; 11: Department of Biological Sciences, Bethel University, St. Paul, MN 55112; 12: Department of Biology, The University of Alabama Birmingham, Birmingham, AL 35294
    AUTHOR AND ARTICLE INFORMATION AUTHOR AND ARTICLE INFORMATION
    • 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].
    Source: J. Microbiol. Biol. Educ. April 2019 vol. 20 no. 1 doi:10.1128/jmbe.v20i1.1702
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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.

References & Citations

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2. Evans Anderson HJ 2017 Abstract: CRISPR in the undergraduate classroom: a CURE FASEB J 31 abstr 589.6
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8. Innovative Genomics Institute 2017 Intensive Undergraduate Summer CRISPR Workshop https://innovativegenomics.org/events/undergrad-workshop-2017/
9. Dahlberg L, Groat Carmona AM 2018 CRISPR-Cas technology in and out of the classroom CRISPR J 1 107 114 10.1089/crispr.2018.0007 http://dx.doi.org/10.1089/crispr.2018.0007
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11. Cooper K, Soneral P, Brownell S 2017 Define your goals before you design a CURE: a call to use backward design in planning course-based undergraduate research experiences J Microbiol Biol Educ 18 2 10.1128/jmbe.v18i2.1287 28861130 5576764 http://dx.doi.org/10.1128/jmbe.v18i2.1287
12. Lave J, Wenger E 1991 Situated learning: legitimate peripheral participation Cambridge University Press Cambridge, UK 10.1017/CBO9780511815355 http://dx.doi.org/10.1017/CBO9780511815355
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15. Courtier-Orgogozo V, Morizot B, Boëte C 2017 Using CRISPR-based gene drive for agriculture pest control EMBO Reports 18 1481 10.15252/embr.201744822 28784604 5579344 http://dx.doi.org/10.15252/embr.201744822
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2019-04-26
2019-08-21

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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Figures

Image of FIGURE 1

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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.

Source: J. Microbiol. Biol. Educ. April 2019 vol. 20 no. 1 doi:10.1128/jmbe.v20i1.1702
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Image of FIGURE 2

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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 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 . CRISPR = clustered regularly interspaced short palindromic repeats; sgRNA = single guide RNA.

Source: J. Microbiol. Biol. Educ. April 2019 vol. 20 no. 1 doi:10.1128/jmbe.v20i1.1702
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Image of FIGURE 3

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FIGURE 3

Sample gels prepared and analyzed by undergraduate students in a summer course demonstrating 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.

Source: J. Microbiol. Biol. Educ. April 2019 vol. 20 no. 1 doi:10.1128/jmbe.v20i1.1702
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Image of FIGURE 4

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FIGURE 4

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

Source: J. Microbiol. Biol. Educ. April 2019 vol. 20 no. 1 doi:10.1128/jmbe.v20i1.1702
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