An Undergraduate Research Project Utilizing CRISPR-Cas9 Gene Editing Technology to Study Gene Function in Arabidopsis thaliana
The CRISPR-Cas9 system functions in microbial viral pathogen recognition pathways by identifying and targeting foreign DNA for degradation. Recently, biotechnological advances have allowed scientists to use CRISPR-Cas9-based elements as a molecular tool to selectively modify DNA in a wide variety o...
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American Society for Microbiology
2019-09-01
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doaj-6ccc23c0eb474d28ab6f92fb40f853882020-11-25T01:16:19ZengAmerican Society for MicrobiologyJournal of Microbiology & Biology Education1935-78771935-78852019-09-0120210.1128/jmbe.v20i2.16661666An Undergraduate Research Project Utilizing CRISPR-Cas9 Gene Editing Technology to Study Gene Function in Arabidopsis thalianaNicholas J. Ruppel0Lauren E. Estell1Robert I. Jackson2Michael J. Wolyniak3Randolph-Macon CollegeRandolph-Macon CollegeHampden-Sydney CollegeHampden-Sydney College The CRISPR-Cas9 system functions in microbial viral pathogen recognition pathways by identifying and targeting foreign DNA for degradation. Recently, biotechnological advances have allowed scientists to use CRISPR-Cas9-based elements as a molecular tool to selectively modify DNA in a wide variety of other living systems. Given the emerging need to bring engaging CRISPR-Cas9 laboratory experiences to an undergraduate audience, we incorporated a CRISPR-based research project into our Genetics class laboratories, emphasizing its use in plants. Our genetic manipulations were designed for Arabidopsis thaliana, which despite serving as a plant research model, has traditionally been difficult to use in a classroom setting. For this project, students transformed plasmid DNA containing the essential CRISPR-Cas9 gene editing elements into A. thaliana. Expression of these elements in the plant genome was expected to create a deletion at one of six targeted genes. The genes we chose had a known seedling and/or juvenile loss-of-function phenotype, which made genetic analysis by students with a limited background possible. It also allowed the project to reach completion in a typical undergraduate semester timeframe. Assessment efforts demonstrated several learning gains, including students’ understanding of CRISPR-Cas9 content, their ability to apply CRISPRCas9 gene editing tools using bioinformatics and genetics, their ability to employ elements of experimental design, and improved science communication skills. They also felt a stronger connection to their scientific education and were more likely to continue on a STEM career path. Overall, this project can be used to introduce CRISPR-Cas9 technology to undergraduates using plants in a single-semester laboratory course. http://jmbesubmissions.asm.org/index.php/jmbe/article/view/1666undergraduate laboratory exerciseCRISPR-Cas9gene editingArabidopsis thalianaplant transformationbacterial transformation |
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DOAJ |
language |
English |
format |
Article |
sources |
DOAJ |
author |
Nicholas J. Ruppel Lauren E. Estell Robert I. Jackson Michael J. Wolyniak |
spellingShingle |
Nicholas J. Ruppel Lauren E. Estell Robert I. Jackson Michael J. Wolyniak An Undergraduate Research Project Utilizing CRISPR-Cas9 Gene Editing Technology to Study Gene Function in Arabidopsis thaliana Journal of Microbiology & Biology Education undergraduate laboratory exercise CRISPR-Cas9 gene editing Arabidopsis thaliana plant transformation bacterial transformation |
author_facet |
Nicholas J. Ruppel Lauren E. Estell Robert I. Jackson Michael J. Wolyniak |
author_sort |
Nicholas J. Ruppel |
title |
An Undergraduate Research Project Utilizing CRISPR-Cas9 Gene Editing Technology to Study Gene Function in Arabidopsis thaliana |
title_short |
An Undergraduate Research Project Utilizing CRISPR-Cas9 Gene Editing Technology to Study Gene Function in Arabidopsis thaliana |
title_full |
An Undergraduate Research Project Utilizing CRISPR-Cas9 Gene Editing Technology to Study Gene Function in Arabidopsis thaliana |
title_fullStr |
An Undergraduate Research Project Utilizing CRISPR-Cas9 Gene Editing Technology to Study Gene Function in Arabidopsis thaliana |
title_full_unstemmed |
An Undergraduate Research Project Utilizing CRISPR-Cas9 Gene Editing Technology to Study Gene Function in Arabidopsis thaliana |
title_sort |
undergraduate research project utilizing crispr-cas9 gene editing technology to study gene function in arabidopsis thaliana |
publisher |
American Society for Microbiology |
series |
Journal of Microbiology & Biology Education |
issn |
1935-7877 1935-7885 |
publishDate |
2019-09-01 |
description |
The CRISPR-Cas9 system functions in microbial viral pathogen recognition pathways by identifying and targeting foreign DNA for degradation. Recently, biotechnological advances have allowed scientists to use CRISPR-Cas9-based elements as a molecular tool to selectively modify DNA in a wide variety of other living systems. Given the emerging need to bring engaging CRISPR-Cas9 laboratory experiences to an undergraduate audience, we incorporated a CRISPR-based research project into our Genetics class laboratories, emphasizing its use in plants. Our genetic manipulations were designed for Arabidopsis thaliana, which despite serving as a plant research model, has traditionally been difficult to use in a classroom setting. For this project, students transformed plasmid DNA containing the essential CRISPR-Cas9 gene editing elements into A. thaliana. Expression of these elements in the plant genome was expected to create a deletion at one of six targeted genes. The genes we chose had a known seedling and/or juvenile loss-of-function phenotype, which made genetic analysis by students with a limited background possible. It also allowed the project to reach completion in a typical undergraduate semester timeframe. Assessment efforts demonstrated several learning gains, including students’ understanding of CRISPR-Cas9 content, their ability to apply CRISPRCas9 gene editing tools using bioinformatics and genetics, their ability to employ elements of experimental design, and improved science communication skills. They also felt a stronger connection to their scientific education and were more likely to continue on a STEM career path. Overall, this project can be used to introduce CRISPR-Cas9 technology to undergraduates using plants in a single-semester laboratory course.
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topic |
undergraduate laboratory exercise CRISPR-Cas9 gene editing Arabidopsis thaliana plant transformation bacterial transformation |
url |
http://jmbesubmissions.asm.org/index.php/jmbe/article/view/1666 |
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