Vivian Nguyen Rose Hills
Engineering Novel Cas9 PAM Targeting Variants for Therapeutic Use
Over the past decade, CRISPR-Cas9 has emerged as an unparalleled tool for single-gene disease therapy. Yet, the vast majority of the genome is currently untargetable by the widely adopted Streptococcus pyogenes Cas9 due to its strict requirement to recognize an NGG protospacer-adjacent motif (PAM) sequence next to the target site. To expand Cas9’s targetable genomic space while maintaining high on-target activity and minimal off-target editing, this research aims to engineer novel SpCas9 variants capable of specifically recognizing non-NGG PAMs through directed evolution in a yeast selection system. In particular, my focus is to re-engineer SpCas9 to specifically target non-canonical NGC>NGT and NGT>NGC PAMs to enable the allele specific editing of a single nucleotide polymorphism (SNP) implicated in Huntington’s Disease. Ultimately, this project will establish a widely applicable methodology to help expand and diversify the therapeutic potential of CRISPR-Cas9.
Message To Sponsor
Thank you for making my summer research possible with your generous sponsorship! I am immensely grateful for this opportunity to continue developing and learning new skills that will be essential as I pursue my goal of attending graduate school. Thank you for your support!