利用光遗传学和 RNAi 鉴定对突触功能重要的基因的多功能学期课程式本科生研究体验。

Eric S Luth, Peter Juo
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摘要

与传统的教学实验活动相比,以课程为基础的本科生研究体验(CURE)可以提高学生的参与度和自信心,提高科学素养,增强批判性思维,促进科学、技术和工程学的普及。在此,我们介绍了针对高年级神经生物学课程的多功能 CURE,该课程将遗传、分子、细胞和行为实验融入到为期一学期的研究中,以确定对 elegans 中谷氨酸突触的形成或功能有重要影响的基因。在介绍了 CURE 方法和 elegans 的基本技术之后,学生们构建了自己的低成本光遗传学装置,通过光刺激激活机械感觉逃逸反射。然后,他们利用这种光激活行为读数进行小规模 RNAi 筛选。一旦确定了感兴趣的基因,学生们就会提交一份研究该基因在神经系统功能中的作用的计划书,并在本学期剩余时间利用突变株开展后续实验。我们还介绍了可以根据教学目标、材料供应情况或教师的研究兴趣对 CURE 进行修改的方法。参加这个实验室大大提高了学生将自己视为 STEM 专业人员的能力,并促使学生报告说,他们在进入研究生院和医学院并取得成功的关键技能方面有了很大的提高。除了 CURE 为学生带来的益处外,教师也从初步数据的生成和学生潜在独立研究项目的培训中获益匪浅。
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A Versatile Semester-Long Course-Based Undergraduate Research Experience using Optogenetics and RNAi to Identify Genes Important for Synapse Function.

Compared to traditional teaching laboratory activities, course-based undergraduate research experiences (CUREs) can increase student engagement and confidence, improve scientific literacy, enhance critical thinking, and promote accessibility in STEM. Here we describe a versatile CURE for an upper-level Neurobiology course that incorporates genetic, molecular, cellular, and behavioral experiments into a semester-long investigation to identify genes important for glutamate synapse formation or function in C. elegans. Following introduction to the CURE approach and basic C. elegans techniques, students construct their own low-cost optogenetics rigs, which we describe in detail here, to activate a mechanosensory escape reflex via photostimulation. They then perform a small-scale RNAi screen with this light-activated behavioral readout. Once a gene of interest is identified, students submit a proposal to investigate the role of this gene in nervous system function and spend the rest of the semester carrying out follow-up experiments using mutant strains. We also describe ways in which this CURE can be modified depending on the pedagogical objectives, availability of materials, or research interests of the instructor. Participating in this lab significantly enhanced students' abilities to see themselves as STEM professionals and prompted students to report substantial gains in skills critical for entry into and success in graduate and medical schools. In addition to the benefits CUREs provide to students, faculty benefit from the generation of preliminary data and training of students for potential independent research projects.

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