A Course-Based Undergraduate Research Experience in CRISPR-Cas9 Experimental Design to Support Reverse Genetic Studies in Arabidopsis thaliana
| Autor: | Jaimie M. Van Norman, David C. Nelson, Berenise Lopez-Lopez, Matthew A. Collin, Susan R. Wessler, Alison M. Mills, James M. Burnette, Venkateswari Jaganatha, Michael Guzman, Carolyn G. Rasmussen, Alejandro Cortez |
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| Rok vydání: | 2021 |
| Předmět: |
0301 basic medicine
Coronavirus disease 2019 (COVID-19) QH301-705.5 Data management education Computational biology General Biochemistry Genetics and Molecular Biology Education 03 medical and health sciences Genome editing Genetics Arabidopsis thaliana CRISPR Biology (General) CURE plant biology LC8-6691 General Immunology and Microbiology biology business.industry 05 social sciences Soft skills 050301 education course-based undergraduate research experience biology.organism_classification Special aspects of education 030104 developmental biology Undergraduate research Curriculum CRISPR-Cas9 remote learning General Agricultural and Biological Sciences business 0503 education Curriculum and Pedagogy Biotechnology Genetic screen |
| Zdroj: | Journal of Microbiology & Biology Education Journal of Microbiology & Biology Education, Vol 22, Iss 2 (2021) Journal of microbiology & biology education, vol 22, iss 2 |
| ISSN: | 1935-7885 1935-7877 |
| DOI: | 10.1128/jmbe.00155-21 |
| Popis: | Gene-editing tools such as CRISPR-Cas9 have created unprecedented opportunities for genetic studies in plants and animals. We designed a course-based undergraduate research experience (CURE) to train introductory biology students in the concepts and implementation of gene-editing technology as well as develop their soft skills in data management and scientific communication. We present two versions of the course that can be implemented with twice-weekly meetings over a 5-week period. In the remote-learning version, students performed homology searches, designed guide RNAs (gRNAs) and primers, and learned the principles of molecular cloning. This version is appropriate when access to laboratory equipment or in-person instruction is limited, such as during closures that have occurred in response to the COVID-19 pandemic. In person, students designed gRNAs, cloned CRISPR-Cas9 constructs, and performed genetic transformation of Arabidopsis thaliana. Students learned how to design effective gRNA pairs targeting their assigned gene with an 86% success rate. Final exams tested students’ ability to apply knowledge of an unfamiliar genome database to characterize gene structure and to properly design gRNAs. Average final exam scores of ∼73% and ∼84% for in-person and remote-learning CUREs, respectively, indicated that students met learning outcomes. The highly parallel nature of the CURE makes it possible to target dozens to hundreds of genes, depending on the number of sections. Applying this approach in a sensitized mutant background enables focused reverse genetic screens for genetic suppressors or enhancers. The course can be adapted readily to other organisms or projects that employ gene editing. |
| Databáze: | OpenAIRE |
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