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你厨房里的CRISPR:一个用于在家编辑酿酒酵母基因的CRISPR试剂盒,用于远程实验室课程。

CRISPR in Your Kitchen: an At-Home CRISPR Kit to Edit Genes in Saccharomyces cerevisiae Used during a Remote Lab Course.

作者信息

McDonnell Lisa, Moore Andrew, Micou Melissa, Day Christopher, Grossman Emily, Meaders Clara

机构信息

Division of Biological Sciences, University of California, San Diego, La Jolla, California, USA.

出版信息

J Microbiol Biol Educ. 2022 Mar 28;23(1). doi: 10.1128/jmbe.00321-21. eCollection 2022 Apr.

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.

摘要

在本科课程中使用基于CRISPR的实验很有吸引力,因为编辑操作简便,且CRISPR与当前研究相关。在新冠疫情之前,我们为一门高注册人数的课程开发了一个面对面的实验室,让学生在酿酒酵母中设计和进行CRISPR编辑实验。疫情之后,实验课程改为线上进行,我们失去了实践环节。随后,我们开发了一个家庭实验套件,其中包含学生培养酿酒酵母并用CRISPR-Cas9诱导编辑所需的DNA分子进行转化所需的所有必要材料。我们的家庭实验套件每个生产成本约为70美元,在2020至2021学年运送给了600多名学生。通过将家庭实验工作添加到我们的远程在线实验课程中,学生能够在ADE2中产生功能缺失突变体(导致红色表型)。学生能够将编辑后的酵母样本送回校园进行测序,从而对CRISPR-Cas9诱导编辑可能产生的不同突变进行表征。在这里,我们描述了生产和使用这些套件的方案,并总结了学生在大型注册远程实验课程中使用家庭实验套件的体验。这些套件为学生在远程课程中参与实践实验提供了机会,也可用于其他领域的学习者,如高中和推广项目。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/d568/9053061/9c248539189f/jmbe.00321-21-f001.jpg

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