A Course-Based Undergraduate Research Experience in CRISPR-Cas9 Experimental Design to Support Reverse Genetic Studies in <italic toggle="yes">Arabidopsis thaliana</italic>

ABSTRACT 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...

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Autores principales:	Alison Mills, Venkateswari Jaganatha, Alejandro Cortez, Michael Guzman, James M. Burnette, Matthew Collin, Berenise Lopez-Lopez, Susan R. Wessler, Jaimie M. Van Norman, David C. Nelson, Carolyn G. Rasmussen
Formato:	article
Lenguaje:	EN
Publicado:	American Society for Microbiology 2021
Materias:	course-based undergraduate research experience CURE remote learning plant biology CRISPR-Cas9 Special aspects of education LC8-6691 Biology (General) QH301-705.5
Acceso en línea:	https://doaj.org/article/237dd0b850654a9c9dfbb40edcdd15b4
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spelling	oai:doaj.org-article:237dd0b850654a9c9dfbb40edcdd15b42021-11-15T15:04:52ZA Course-Based Undergraduate Research Experience in CRISPR-Cas9 Experimental Design to Support Reverse Genetic Studies in <italic toggle="yes">Arabidopsis thaliana</italic>10.1128/jmbe.00155-211935-78851935-7877https://doaj.org/article/237dd0b850654a9c9dfbb40edcdd15b42021-09-01T00:00:00Zhttps://journals.asm.org/doi/10.1128/jmbe.00155-21https://doaj.org/toc/1935-7877https://doaj.org/toc/1935-7885ABSTRACT 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.Alison MillsVenkateswari JaganathaAlejandro CortezMichael GuzmanJames M. BurnetteMatthew CollinBerenise Lopez-LopezSusan R. WesslerJaimie M. Van NormanDavid C. NelsonCarolyn G. RasmussenAmerican Society for Microbiologyarticlecourse-based undergraduate research experienceCUREremote learningplant biologyCRISPR-Cas9Special aspects of educationLC8-6691Biology (General)QH301-705.5ENJournal of Microbiology & Biology Education, Vol 22, Iss 2 (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	course-based undergraduate research experience CURE remote learning plant biology CRISPR-Cas9 Special aspects of education LC8-6691 Biology (General) QH301-705.5
spellingShingle	course-based undergraduate research experience CURE remote learning plant biology CRISPR-Cas9 Special aspects of education LC8-6691 Biology (General) QH301-705.5 Alison Mills Venkateswari Jaganatha Alejandro Cortez Michael Guzman James M. Burnette Matthew Collin Berenise Lopez-Lopez Susan R. Wessler Jaimie M. Van Norman David C. Nelson Carolyn G. Rasmussen A Course-Based Undergraduate Research Experience in CRISPR-Cas9 Experimental Design to Support Reverse Genetic Studies in <italic toggle="yes">Arabidopsis thaliana</italic>
description	ABSTRACT 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.
format	article
author	Alison Mills Venkateswari Jaganatha Alejandro Cortez Michael Guzman James M. Burnette Matthew Collin Berenise Lopez-Lopez Susan R. Wessler Jaimie M. Van Norman David C. Nelson Carolyn G. Rasmussen
author_facet	Alison Mills Venkateswari Jaganatha Alejandro Cortez Michael Guzman James M. Burnette Matthew Collin Berenise Lopez-Lopez Susan R. Wessler Jaimie M. Van Norman David C. Nelson Carolyn G. Rasmussen
author_sort	Alison Mills
title	A Course-Based Undergraduate Research Experience in CRISPR-Cas9 Experimental Design to Support Reverse Genetic Studies in <italic toggle="yes">Arabidopsis thaliana</italic>
title_short	A Course-Based Undergraduate Research Experience in CRISPR-Cas9 Experimental Design to Support Reverse Genetic Studies in <italic toggle="yes">Arabidopsis thaliana</italic>
title_full	A Course-Based Undergraduate Research Experience in CRISPR-Cas9 Experimental Design to Support Reverse Genetic Studies in <italic toggle="yes">Arabidopsis thaliana</italic>
title_fullStr	A Course-Based Undergraduate Research Experience in CRISPR-Cas9 Experimental Design to Support Reverse Genetic Studies in <italic toggle="yes">Arabidopsis thaliana</italic>
title_full_unstemmed	A Course-Based Undergraduate Research Experience in CRISPR-Cas9 Experimental Design to Support Reverse Genetic Studies in <italic toggle="yes">Arabidopsis thaliana</italic>
title_sort	course-based undergraduate research experience in crispr-cas9 experimental design to support reverse genetic studies in <italic toggle="yes">arabidopsis thaliana</italic>
publisher	American Society for Microbiology
publishDate	2021
url	https://doaj.org/article/237dd0b850654a9c9dfbb40edcdd15b4
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A Course-Based Undergraduate Research Experience in CRISPR-Cas9 Experimental Design to Support Reverse Genetic Studies in <italic toggle="yes">Arabidopsis thaliana</italic>

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