Lipid Nanoparticle-Mediated Delivery of CRISPR-Cas9 for Therapeutic Gene Correction

Abstract

The thesis “Lipid Nanoparticle-Mediated Delivery of CRISPR-Cas9 for Therapeutic Gene Correction” delves into the potential of CRISPR-Cas9 as a gene therapy tool for treating hereditary disorders. Originally a bacterial defense mechanism, CRISPR-Cas9 has been modified into a precision gene editing tool. It functions like molecular scissors, creating specific DNA breaks that trigger DNA repair processes, which, when coupled with repair DNA, can correct gene mutations causing hereditary diseases. However, the challenge lies in safely and efficiently delivering CRISPR-Cas9 to affected organs in patients.

The thesis explores the use of lipid nanoparticles, small, virus-sized particles composed of naturally occurring body fats, as a viable delivery system for CRISPR-Cas9. The work in this thesis demonstrates that the designed lipid nanoparticles can effectively transport CRISPR-Cas9 into both cultured cells and later in liver cells of mice as well. Moreover, the results show that the delivered CRISPR-Cas9 generates precise genome edits. Furthermore, it addresses the potential immune responses against CRISPR-Cas9, given it bacterial origin, and introduces a strategy to mitigate such reactions.

In summary, this thesis presents a promising approach to deliver CRISPR-Cas9 to diseased cells, enabling the precise correction of pathogenic gene mutations, marking a significant advancement in the field of gene therapy for hereditary disorders.