An Undergraduate Research Project Utilizing CRISPR-Cas9 Gene Editing Technology to Study Gene Function in <italic toggle="yes">Arabidopsis thaliana</italic>

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...

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Autores principales:	Nicholas J. Ruppel, Lauren E. Estell, Robert I. Jackson, Michael J. Wolyniak
Formato:	article
Lenguaje:	EN
Publicado:	American Society for Microbiology 2019
Materias:	Special aspects of education LC8-6691 Biology (General) QH301-705.5
Acceso en línea:	https://doaj.org/article/23731ca3f1204e7ea2759971f55f6166
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spelling	oai:doaj.org-article:23731ca3f1204e7ea2759971f55f61662021-11-15T15:04:28ZAn Undergraduate Research Project Utilizing CRISPR-Cas9 Gene Editing Technology to Study Gene Function in <italic toggle="yes">Arabidopsis thaliana</italic>10.1128/jmbe.v20i2.16661935-78851935-7877https://doaj.org/article/23731ca3f1204e7ea2759971f55f61662019-01-01T00:00:00Zhttps://journals.asm.org/doi/10.1128/jmbe.v20i2.1666https://doaj.org/toc/1935-7877https://doaj.org/toc/1935-7885The 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 CRISPR-Cas9 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.Nicholas J. RuppelLauren E. EstellRobert I. JacksonMichael J. WolyniakAmerican Society for MicrobiologyarticleSpecial aspects of educationLC8-6691Biology (General)QH301-705.5ENJournal of Microbiology & Biology Education, Vol 20, Iss 2 (2019)
institution	DOAJ
collection	DOAJ
language	EN
topic	Special aspects of education LC8-6691 Biology (General) QH301-705.5
spellingShingle	Special aspects of education LC8-6691 Biology (General) QH301-705.5 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 <italic toggle="yes">Arabidopsis thaliana</italic>
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 CRISPR-Cas9 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.
format	article
author	Nicholas J. Ruppel Lauren E. Estell Robert I. Jackson Michael J. Wolyniak
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 <italic toggle="yes">Arabidopsis thaliana</italic>
title_short	An Undergraduate Research Project Utilizing CRISPR-Cas9 Gene Editing Technology to Study Gene Function in <italic toggle="yes">Arabidopsis thaliana</italic>
title_full	An Undergraduate Research Project Utilizing CRISPR-Cas9 Gene Editing Technology to Study Gene Function in <italic toggle="yes">Arabidopsis thaliana</italic>
title_fullStr	An Undergraduate Research Project Utilizing CRISPR-Cas9 Gene Editing Technology to Study Gene Function in <italic toggle="yes">Arabidopsis thaliana</italic>
title_full_unstemmed	An Undergraduate Research Project Utilizing CRISPR-Cas9 Gene Editing Technology to Study Gene Function in <italic toggle="yes">Arabidopsis thaliana</italic>
title_sort	undergraduate research project utilizing crispr-cas9 gene editing technology to study gene function in <italic toggle="yes">arabidopsis thaliana</italic>
publisher	American Society for Microbiology
publishDate	2019
url	https://doaj.org/article/23731ca3f1204e7ea2759971f55f6166
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An Undergraduate Research Project Utilizing CRISPR-Cas9 Gene Editing Technology to Study Gene Function in <italic toggle="yes">Arabidopsis thaliana</italic>

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