Various CRISPR-Cas9-derived tools enable genome engineering, allowing efficient gene knock-out, knock-in, and site-specific regulation of epigenetic events. The course will provide hands-on training in genome editing (the prototypic knock out) using the first generation of CRISPR-Cas9 tools (Cas9 and sgRNA expressing plasmids). In a second experiment students will be using sgRNA directing recombinant dCas9 to modulate gene expression. We will discuss the importance of later editing generations using Cas9 RNA, and the Cas9 protein, along with an appropriate single-guide RNA (sgRNA) to knock-out, knock-in, and edit at specific genomic loci.
To accompany the practical part, we will discuss various relevant topics, and to name just a few, we will study the emergence of several CRISPR-derived technologies as a vital discipline relevant for research and biotechnology in cells and organisms alike. This includes the current, yet limited, knowledge of the cellular DNA repair machineries that underlie efficient genomic modifications and the basic knowledge required for efficient gene knock-out and knock-in that should be characterized by minimal undesired “off-targets”.