An Undergraduate Research Project Utilizing CRISPR-Cas9 Gene Editing Technology to Study Gene Function in Arabidopsis thaliana†
An Undergraduate Research Project Utilizing CRISPR-Cas9 Gene Editing Technology to Study Gene Function in Arabidopsis thaliana†
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利用 CRISPR-Cas9 基因编辑技术研究拟南芥基因功能的本科研究项目†
DOI：
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发表时间：
2019
影响因子：
1.9
通讯作者：
M. Wolyniak
中科院分区：
文献类型：
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作者：
N. Ruppel;Lauren E. Estell;R. Jackson;M. Wolyniak
CRISPR-Cas9系统通过识别和靶向外源DNA进行降解，在微生物病毒病原体识别途径中发挥作用。最近，生物技术的进步使科学家们能够使用基于CRISPR-Cas9的元件作为分子工具，在各种其他生命系统中选择性地修饰DNA。考虑到将CRISPR-Cas9实验室体验带给本科生的需求，我们将一个基于CRISPR的研究项目纳入了我们的遗传学类实验室，强调其在植物中的应用。我们的遗传操作是为拟南芥设计的，尽管它是一种植物研究模型，但传统上很难在课堂上使用。在这个项目中，学生们将含有基本CRISPR-Cas9基因编辑元件的质粒DNA转化为A. thaliana.这些元件在植物基因组中的表达预期在六个靶基因之一处产生缺失。我们选择的基因有一个已知的幼苗和/或少年功能丧失表型，这使得遗传分析的学生与有限的背景可能。它还允许该项目在典型的本科学期时间内完成。评估工作显示了几项学习成果，包括学生对CRISPR-Cas9内容的理解，他们使用生物信息学和遗传学应用CRISPR-Cas9基因编辑工具的能力，他们使用实验设计元素的能力，以及提高的科学交流技能。他们也觉得自己与科学教育有更强的联系，更有可能继续走STEM职业道路。总的来说，这个项目可以用来在一个学期的实验室课程中向本科生介绍CRISPR-Cas9技术。
The CRISPR-Cas9 system functions in microbial viral pathogen recognition pathways by identifying and targeting foreign DNA for degradation. Recently, biotechnological advances have allowed scientists to use CRISPR-Cas9-based elements as a molecular tool to selectively modify DNA in a wide variety of other living systems. Given the emerging need to bring engaging CRISPR-Cas9 laboratory experiences to an undergraduate audience, we incorporated a CRISPR-based research project into our Genetics class laboratories, emphasizing its use in plants. Our genetic manipulations were designed for Arabidopsis thaliana, which despite serving as a plant research model, has traditionally been difficult to use in a classroom setting. For this project, students transformed plasmid DNA containing the essential CRISPR-Cas9 gene editing elements into A. thaliana. Expression of these elements in the plant genome was expected to create a deletion at one of six targeted genes. The genes we chose had a known seedling and/or juvenile loss-of-function phenotype, which made genetic analysis by students with a limited background possible. It also allowed the project to reach completion in a typical undergraduate semester timeframe. Assessment efforts demonstrated several learning gains, including students’ understanding of CRISPR-Cas9 content, their ability to apply CRISPR-Cas9 gene editing tools using bioinformatics and genetics, their ability to employ elements of experimental design, and improved science communication skills. They also felt a stronger connection to their scientific education and were more likely to continue on a STEM career path. Overall, this project can be used to introduce CRISPR-Cas9 technology to undergraduates using plants in a single-semester laboratory course.
影响因子：
5.6
作者：
Brokowski C;Adli M
通讯作者：
Adli M