Andrew Chen L&S Sciences
Optimization of CRISPR-Cas9 using an Adeno-Associated Virus (AAV) Delivered Self-Inactivation System
CRISPR-Cas9 is a versatile genome-editing technology that is widely used to modify genome sequences linked to genetic abnormalities. Often, the CRISPR-Cas9 system is used to target or repair disease-causing versions of genes, delete fragments of unwanted DNA, and insert therapeutic gene sequences to treat diseases. However, many studies have shown that CRISPR-Cas9 can cause unexpected mutations and genomic rearrangements in untargeted sites. My research aims to develop a self-inactivation system in effort to limit long-term expression of nuclease-editing protein Cas9 while still maximizing its on-target cuts and reducing off-target effects. To effectively deliver the Cas9-sgRNA complex, I plan to use adeno-associated virus (AAV) due to its highly efficient, targeted gene delivery. In addition, I plan to modulate the kinetics of Cas9 targeting the single-guide RNA to gain better control of Cas9 activity for potential in vivo applications. The goal of this work is to improve current CRISPR-Cas9 applications in not only labs but also future clinical applications in which AAV delivery may be common for gene therapy.