Development of a Freeze-Dried CRISPR-Cas12 Sensor for Detecting Wolbachia in the Secondary Science Classroom
Development of a Freeze-Dried CRISPR-Cas12 Sensor for Detecting Wolbachia in the Secondary Science Classroom
复制标题
DOI：
10.1021/acssynbio.1c00503
复制
发表时间：
2022-02-18
影响因子：
4.7
通讯作者：
Jewett, Michael C.
中科院分区：
文献类型：
--
作者：
Rybnicky, Grant A.;Dixon, Radeen A.;Jewett, Michael C.
培训未来的合成生物学劳动力需要让学生有机会在中学教育中接触生物技术概念和活动。利用聚合酶链式反应 (PCR) 检测节肢动物中的沃尔巴克氏体细菌已成为中学生在科学课堂上研究和应用重组 DNA 技术的常见方法。尽管这是一项重要的活动，但尖端生物技术（例如基于聚类规则间隔短回文重复 (CRISPR) 的诊断）尚未在课堂上广泛实施。为了弥补这一差距，我们提出了一种冻干 CRISPR-Cas12 传感反应，以补充传统的重组 DNA 技术教育并教授合成生物学概念。该反应可在 2 小时内从节肢动物来源的 PCR 样本中准确检测出沃尔巴克氏体，并且可以在室温下保存一个多月而不会明显降解。这些反应易于使用，对于 22 名学生的教室来说，实施成本不到 40 美元，其中包括可重复使用设备的成本。我们认为这些冷冻干燥的 CRISPR-Cas12 反应是将合成生物学教育纳入现有生物学课程的一种可行方式，这将扩大科学、技术、工程和数学方面的生物学教育机会。
Training the future synthetic biology workforce requires the opportunity for students to be exposed to biotechnology concepts and activities in secondary education. Detecting Wolbachia bacteria in arthropods using polymerase chain reaction (PCR) has become a common way for secondary students to investigate and apply recombinant DNA technology in the science classroom. Despite this important activity, cutting-edge biotechnologies such as clustered regularly interspaced short palindromic repeat (CRISPR)-based diagnostics have yet to be widely implemented in the classroom. To address this gap, we present a freeze-dried CRISPR-Cas12 sensing reaction to complement traditional recombinant DNA technology education and teach synthetic biology concepts. The reactions accurately detect Wolbachia from arthropod-derived PCR samples in under 2 h and can be stored at room temperature for over a month without appreciable degradation. The reactions are easy-to-use and cost less than $40 to implement for a classroom of 22 students including the cost of reusable equipment. We see these freezedried CRISPR-Cas12 reactions as an accessible way to incorporate synthetic biology education into the existing biology curriculum, which will expand biology educational opportunities in science, technology, engineering, and mathematics.