Lab Course: Mammalian Gene Editing

Abstract

The students are trained in the basic techniques of designing, constructing, conducting, and characterizing CRISPR-Cas gene editing experiments in mammalian cells. CRISPR-Cas plasmids are cloned with custom guide RNAs targeting genes of interest followed by transient transfection into mammalian cells and the analysis of gene editing efficiency.

Objective

The objective of the course is to design and conduct a gene editing experiment, including selecting the target gene, designing the CRISPR-Cas9 reagents, cloning expression plasmids, delivering plasmids into mammalian cells, and analyzing gene editing outcomes.

Content

The course will take place over 5 weeks, with 2 days per week spent on lab work. The 4 weeks will be dedicated to the following activities Week 1: Introduction to the course; supervised design of gRNAs and gene editing analysis reagents as well as detailed planning. Week 2: Cloning of the constructs part 1; optimization of gene editing analysis reagents. Week 3: Cloning of the constructs part 2; mammalian cell transfection. Week 4: Gene editing analysis. Week 5: Data analysis and preparation of the final report.

Resources