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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.

DOI:10.1128/jmbe.00321-21
PMID:35496692
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC9053061/
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/d568/9053061/4930b152d879/jmbe.00321-21-f002.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/d568/9053061/9c248539189f/jmbe.00321-21-f001.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/d568/9053061/4930b152d879/jmbe.00321-21-f002.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/d568/9053061/9c248539189f/jmbe.00321-21-f001.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/d568/9053061/4930b152d879/jmbe.00321-21-f002.jpg

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本文引用的文献

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CRISPR-Cas9 Gene Editing in Yeast: A Molecular Biology and Bioinformatics Laboratory Module for Undergraduate and High School Students.酵母中的CRISPR-Cas9基因编辑:面向本科生和高中生的分子生物学与生物信息学实验室模块
J Microbiol Biol Educ. 2021 May 31;22(2). doi: 10.1128/jmbe.00106-21. eCollection 2021 Fall.
2
CRISPR Gene Editing in Yeast: An Experimental Protocol for an Upper-Division Undergraduate Laboratory Course.酵母中的CRISPR基因编辑:一门本科高年级实验课程的实验方案
Biochem Mol Biol Educ. 2018 Nov;46(6):592-601. doi: 10.1002/bmb.21175. Epub 2018 Oct 12.
3
Genome engineering in Saccharomyces cerevisiae using CRISPR-Cas systems.
J Microbiol Biol Educ. 2024 Aug 29;25(2):e0018723. doi: 10.1128/jmbe.00187-23. Epub 2024 May 10.
4
Are null segregants new combinations of heritable material and should they be regulated?零分离体是可遗传物质的新组合吗?它们应该受到监管吗?
Front Genome Ed. 2023 Jan 10;4:1064103. doi: 10.3389/fgeed.2022.1064103. eCollection 2022.
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Cloud-controlled microscopy enables remote project-based biology education in underserved Latinx communities.云控显微镜助力在服务不足的拉丁裔社区开展基于项目的远程生物教育。
Heliyon. 2022 Nov 15;8(11):e11596. doi: 10.1016/j.heliyon.2022.e11596. eCollection 2022 Nov.
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Opportunities and Challenges of Online Instruction and Effective Pedagogy That Blurs the Lines between Online and On-Site Teaching and Learning.在线教学的机遇与挑战以及模糊线上与线下教学界限的有效教学法
J Microbiol Biol Educ. 2022 Apr 11;23(1). doi: 10.1128/jmbe.00047-22. eCollection 2022 Apr.
利用 CRISPR-Cas 系统进行酿酒酵母的基因组工程。
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