HTGAA Class 7: CRISPR editing

​Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR) is a gene-editing tool that allows scientists to modify an organism’s DNA. The system is based on a naturally occurring defense mechanism found in bacteria to fight off viral invaders by cutting their DNA.

​The CRISPR-Cas9 system consists of two main components: the Cas9 protein, which acts as a molecular scissor, and a guide RNA (gRNA) that directs the Cas9 protein to a specific target sequence in the DNA. When the Cas9 protein and the gRNA are introduced into a cell, the gRNA binds to the target DNA sequence, guiding the Cas9 protein to make a precise cut in the DNA, technically referred to as DNA double-strand break (DSB) later. Once the DNA is cut, the cell’s own repair machinery attempts to fix the break, which can result in the inactivation of a gene.

​In this laboratory, we will explore the cutting-edge CRISPR-prime editing system, a powerful tool for genetic engineering that enables insertions, deletions, and substitutions in target DNA sequences without introducing double-strand breaks. Particularly, this experiment will demonstrate how to practically perform gene deletion using CRISPR editing in E. coli which can be broadly applied to other designs for various applications. This system has broad applications in both basic research and biotechnology, including gene therapy, metabolic engineering, and the development of novel organisms with unique characteristics.