摘要
CRISPR/Cas9系统作为一项开创性的基因编辑技术,已被广泛地应用于各种生物体的基因研究,并将彻底改变生物医学研究以及疾病治疗。理解与掌握CRISPR技术的基本原理和操作方法,能够帮助生物相关专业学生紧跟时代前沿技术,培养创新科研思维。该研究设计了基于模式真菌酿酒酵母(Saccharomyces cerevisiae)的CRISPR/Cas9基因编辑教学实验,并融合了GFP荧光标记技术增强实验教学的可视化。实验模块包括gRNA设计、pCAS/gRNA共表达质粒构建、同源修复DNA表达盒构建、PCR产物纯化、醋酸锂酵母转化和荧光显微镜观察GFP表达等。教学实践表明学生能完成酿酒酵母CRISPR基因编辑操作模块,得到预期结果。该团队在实验教学过程中充分利用超星学习通平台进行多元智能评价,教学评价显示学生更加了解CRISPR基因编辑技术在生物研究中的重要性,并显著提高了本科生从事科学研究的兴趣。在新工科教育背景下,将此前沿技术引入本科生实验教学,有助于提高教学质量。
As an emerging gene editing technology,the CRISPR/Cas9 system is widely used in the research of various organisms and is bound to revolutionize biomedical research and disease treatment.Understanding and mastering CRISPR technology will provide a strong foundation for undergraduates majoring in biology and related fields to follow the advanced technology,and cultivate innovative thinking.This study developed an innovative teaching experiment to apply CRISPR/Cas9 technology in Saccharomyces cerevisiae and incorporate GFP fluorescent labeling technique into this gene editing experiment to improve the visualization.The experimental modules include the design of gRNA,the construction of pCAS/gRNA coexpression plasmids,the construction of DNA cassette for homologous repairing,PCR product purification,lithium acetate-mediated yeast transformation and fluorescence microscope observation of GFP expression.Teaching practice indicates that students could complete the laboratory modules and achieve the expected outcomes.Multiple assessments were performed using intelligent evaluation platform throughout the entire laboratory process.Post-class assessment indicates that this team’s experimental teaching course allows undergraduates to deepen their understanding of CRISPR/Cas9 technology and increase their interest in scientific research.Under the background of emerging engineering education,it is of great significance to introduce this cutting-edge scientific technique into the undergraduate classroom to improve the teaching quality.
作者
王文洁
杨玲琦
李余动
WANG Wenjie;YANG Lingqi;LI Yudong(School of Food Science and Biotechnology,Zhejiang Gongshang University,Hangzhou 310018,China)
出处
《中国细胞生物学学报》
2025年第7期1668-1675,共8页
Chinese Journal of Cell Biology
基金
浙江省教育厅一般科研项目(批准号:Y202250153)
浙江省“十四五”研究生教育改革项目(批准号:SYJG202201006)资助的课题。
