In silico optimization of CRISPR/Cas9-Gold nanovehicle design

CRISPR/Cas9 gene editing is a novel and versatile technology that correct disease-causing errors in the genomes, with multiple and broad applications in medical, agricultural and pharmaceutical industries. The major obstacle in implementing CRISPR/Cas9-based therapeutics is to develop nanovehicles that can target genetic mutations in a specific manner. Gold nanoparticles (GNP) functionalized with synthetic oligonucleotides combine the optoelectronic properties of the gold core with the particular properties of oligonucleotides, for instance selectivity and specificity. Furthermore, functionalized GNPs have been previously used in vivo for delivery of CRISPR/Cas9 to successfully repair gene mutations causing muscular dystrophy. However, prior to clinical applications, safe and efficient CRISPR/Cas9-Gold-based delivery vehicles should be designed based on a deep understanding of the underlying molecular mechanisms. In this proposal, we intend to computationally optimize such a design in two steps. First, we will optimize the DNA loading on a range of GNP sizes in nanoparticle-oligonucleotide conjugates. In the second stage, we will perform a thorough in silico investigation of GNP-oligonucleotide conjugates complexed with Cas9 ribonucleoprotein as delivery nanovehicles.