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CRISPR in Your Kitchen: an At-Home CRISPR Kit to Edit Genes in Saccharomyces Cerevisiae Used During a Remote Lab Course

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Specialty Biology
Date 2022 May 2
PMID 35496692
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Abstract

The use of CRISPR-based experiments in an undergraduate course is appealing because of the ease of editing, and the relevance of CRISPR to current research. Before the COVID-19 pandemic, we developed an in-person lab for a high-enrollment course that allowed students to design and conduct CRISPR editing experiments in budding yeast, Saccharomyces cerevisiae. Post pandemic, the lab course moved online, and we lost the hands-on component. We subsequently developed an at-home kit that contained all the necessary materials for students to grow and transform S. cerevisiae with the DNA molecules necessary for CRISPR-Cas9 induced editing. Our at-home kits cost approximately $70 each to produce and were shipped to over 600 students during the 2020 to 2021 academic year. By adding the at-home experimental work to our remote, online lab course, students were able to generate loss-of-function mutants in ADE2 (causing a red color phenotype). Students were able to send edited yeast samples back to campus for sequencing, allowing for characterization of the different mutations that can occur due to CRISPR-Cas9 induced editing. Here, we described the protocol to produce and use the kits and summarized the student experience of using the at-home kit in a large enrollment, remote lab course. These kits provided opportunities to engage students in hands-on experimentation during a remote course and could also be used to reach learners in other domains, such as high schools and outreach programs.

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References
1.
Ens S, Olson A, Dudley C, Ross 3rd N, Siddiqi A, Umoh K . Inexpensive and safe DNA gel electrophoresis using household materials. Biochem Mol Biol Educ. 2012; 40(3):198-203. DOI: 10.1002/bmb.20596. View

2.
DiCarlo J, Norville J, Mali P, Rios X, Aach J, Church G . Genome engineering in Saccharomyces cerevisiae using CRISPR-Cas systems. Nucleic Acids Res. 2013; 41(7):4336-43. PMC: 3627607. DOI: 10.1093/nar/gkt135. View

3.
Sehgal N, Sylves M, Sahoo A, Chow J, Walker S, Cullen P . CRISPR Gene Editing in Yeast: An Experimental Protocol for an Upper-Division Undergraduate Laboratory Course. Biochem Mol Biol Educ. 2018; 46(6):592-601. PMC: 6615721. DOI: 10.1002/bmb.21175. View

4.
Sankaran S, Smith J, Roy K . CRISPR-Cas9 Gene Editing in Yeast: A Molecular Biology and Bioinformatics Laboratory Module for Undergraduate and High School Students. J Microbiol Biol Educ. 2021; 22(2). PMC: 8442027. DOI: 10.1128/jmbe.00106-21. View