CRISPR/Cas9 Mechanism and Applications Progression

Abstract

CRISPR (short for Clustered Regularly Interspaced Short Palindromic Repeats) and the accompanying endonuclease Cas9 are potent tools for gene editing in many applied disciplines. The CRISPR/Cas9 system primarily functions through two components, in which the single guide RNA (sgRNA) directs the Cas9 protein with endonuclease activity to carry out precise DNA cleavage. The cell's inherent repair mechanisms allow it to complete the subsequent reconnection of the broken genes. Gene knock-in or knock-out can therefore be achieved via this approach. This system was typically applied to prokaryotes in early studies. Its role as a component of their defense mechanism has been thoroughly studied. With rapid development over the past ten years, this system can now be effectively used in a multitude of eukaryotic cells, including yeast cells and mammalian cells. Because of its accuracy, effectiveness, and low cost, CRISPR/Cas9 has been widely used for human disease treatment and has shown great potential in gene therapies for hereditary diseases. However, since more late-stage clinical trials are required to validate its safety, its applications still face many challenges. This review investigates the mechanism of CRISPR/Cas9-mediated gene editing technology and highlights its recent applications in Alzheimer's disease, Sickle cell disease, and Duchenne muscular dystrophy, which are difficult to treat with current approaches. Some challenges the CRISPR/Cas9 system has encountered at this stage are described, and its prospects are also examined.