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一段关于CRISPR-Cas9介导的真核生物基因组编辑的本科研究经历,用于教授基础生物化学技术。

An undergraduate research experience in CRISPR-Cas9 mediated eukaryotic genome editing to teach fundamental biochemistry techniques.

作者信息

Tonsager Andrew J, Stargell Laurie A

机构信息

Department of Biochemistry and Molecular Biology, Colorado State University, Fort Collins, Colorado, USA.

出版信息

Biochem Mol Biol Educ. 2025 Jan-Feb;53(1):33-45. doi: 10.1002/bmb.21862. Epub 2024 Oct 8.

Abstract

CRISPR-Cas9 technology is an established, powerful tool for genome editing through the ability to target specific DNA sequences of interest for introduction of desired genetic modifications. CRISPR-Cas9 is utilized for a variety of purposes, ranging from a research molecular biology tool to treatment for human diseases. Due to its prominence across a variety of applications, it is critical that undergraduates in the life sciences are educated on CRISPR-Cas9 technology. To this end, we created an intensive eight-week long course-based undergraduate research experience (CURE) designed for students to understand CRISPR-Cas9 genome editing and perform it in Saccharomyces cerevisiae. Students enrolled in the CURE participate in 2, 3-h sessions a week and are engaged in the entire process of CRISPR-Cas9 genome editing, from preparation of genome editing reagents to characterization of mutant yeast strains. During the process, students master fundamental techniques in the life sciences, including sterile technique, Polymerase Chain Reaction (PCR), primer design, sequencing requirements, and data analysis. The course is developed with flexibility in the schedule for repetition of techniques in the event of a failed experiment, providing an authentic research experience for the students. Additionally, we have developed the course to be easily modified for the editing of any yeast gene, offering the potential to expand the course in research-driven classroom or laboratory settings.

摘要

CRISPR-Cas9技术是一种成熟且强大的基因组编辑工具,它能够靶向特定的目标DNA序列,以引入所需的基因修饰。CRISPR-Cas9有多种用途,从研究分子生物学工具到人类疾病治疗。鉴于其在各种应用中的突出地位,对生命科学专业的本科生进行CRISPR-Cas9技术教育至关重要。为此,我们创建了一个为期八周的强化课程式本科研究体验(CURE),旨在让学生了解CRISPR-Cas9基因组编辑并在酿酒酵母中进行操作。参加CURE的学生每周参加两次3小时的课程,并参与CRISPR-Cas9基因组编辑的全过程,从基因组编辑试剂的制备到突变酵母菌株的鉴定。在此过程中,学生掌握生命科学的基本技术,包括无菌技术、聚合酶链反应(PCR)、引物设计、测序要求和数据分析。该课程在实验失败时可灵活安排技术重复的时间,为学生提供真实的研究体验。此外,我们开发的课程易于修改以编辑任何酵母基因,为在以研究为导向的课堂或实验室环境中扩展课程提供了可能性。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/0297/11752412/10bd90802c42/BMB-53-33-g003.jpg

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