An Introduction to CRISPR-Mediated Genome Editing in Fungi

Central dogma, transformation, and genome editing are key biological concepts for which junior scientists must gain experience during training. Here we present an exercise that introduces these concepts in a single practical laboratory exercise. Our exercise utilizes CRISPR/Cas9 genome editing to in...

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Autores principales:	Valmik K. Vyas, Douglas A. Bernstein
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/49608ca699394ec9a28a316c0e36aa64
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spelling	oai:doaj.org-article:49608ca699394ec9a28a316c0e36aa642021-11-15T15:04:12ZAn Introduction to CRISPR-Mediated Genome Editing in Fungi10.1128/jmbe.v20i3.18091935-78851935-7877https://doaj.org/article/49608ca699394ec9a28a316c0e36aa642019-01-01T00:00:00Zhttps://journals.asm.org/doi/10.1128/jmbe.v20i3.1809https://doaj.org/toc/1935-7877https://doaj.org/toc/1935-7885Central dogma, transformation, and genome editing are key biological concepts for which junior scientists must gain experience during training. Here we present an exercise that introduces these concepts in a single practical laboratory exercise. Our exercise utilizes CRISPR/Cas9 genome editing to introduce a stop codon into Saccharomyces cerevisiae ADE2. This edit leads to the buildup of an adenine precursor that dyes the edited cells red. As the repair template, guide RNA, and Cas9 are all encoded in our vector, transformation can be performed in 2 hours. Furthermore, since all components of the Cas9/CRISPR system are encoded by the vector, specialized equipment and reagents, such as a PCR machine or oligonucleotides, are not required to perform the experiments as designed. As such, these exercises provide an efficient cost-effective introduction to a wide variety of key molecular biology concepts and lay the foundation for more rigorous investigations in upper-level classes and independent research projects.Valmik K. VyasDouglas A. BernsteinAmerican Society for MicrobiologyarticleSpecial aspects of educationLC8-6691Biology (General)QH301-705.5ENJournal of Microbiology & Biology Education, Vol 20, Iss 3 (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 Valmik K. Vyas Douglas A. Bernstein An Introduction to CRISPR-Mediated Genome Editing in Fungi
description	Central dogma, transformation, and genome editing are key biological concepts for which junior scientists must gain experience during training. Here we present an exercise that introduces these concepts in a single practical laboratory exercise. Our exercise utilizes CRISPR/Cas9 genome editing to introduce a stop codon into Saccharomyces cerevisiae ADE2. This edit leads to the buildup of an adenine precursor that dyes the edited cells red. As the repair template, guide RNA, and Cas9 are all encoded in our vector, transformation can be performed in 2 hours. Furthermore, since all components of the Cas9/CRISPR system are encoded by the vector, specialized equipment and reagents, such as a PCR machine or oligonucleotides, are not required to perform the experiments as designed. As such, these exercises provide an efficient cost-effective introduction to a wide variety of key molecular biology concepts and lay the foundation for more rigorous investigations in upper-level classes and independent research projects.
format	article
author	Valmik K. Vyas Douglas A. Bernstein
author_facet	Valmik K. Vyas Douglas A. Bernstein
author_sort	Valmik K. Vyas
title	An Introduction to CRISPR-Mediated Genome Editing in Fungi
title_short	An Introduction to CRISPR-Mediated Genome Editing in Fungi
title_full	An Introduction to CRISPR-Mediated Genome Editing in Fungi
title_fullStr	An Introduction to CRISPR-Mediated Genome Editing in Fungi
title_full_unstemmed	An Introduction to CRISPR-Mediated Genome Editing in Fungi
title_sort	introduction to crispr-mediated genome editing in fungi
publisher	American Society for Microbiology
publishDate	2019
url	https://doaj.org/article/49608ca699394ec9a28a316c0e36aa64
work_keys_str_mv	AT valmikkvyas anintroductiontocrisprmediatedgenomeeditinginfungi AT douglasabernstein anintroductiontocrisprmediatedgenomeeditinginfungi AT valmikkvyas introductiontocrisprmediatedgenomeeditinginfungi AT douglasabernstein introductiontocrisprmediatedgenomeeditinginfungi
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An Introduction to CRISPR-Mediated Genome Editing in Fungi

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