(Invited) CRISPR-Cas9-Based Cancer Therapy Delivery By Carbon Nanomaterials

Abstract

Gene therapeutics is a rapidly developing field promising critical advances in finding new vaccines and treatments, as well as genetic adjustments to plants and animal species. This novel area of science may generate treatment pathways for complex diseases such as cancer, Alzheimer’s or sickle cell anemia that can not be easily treated by conventional therapies. Recently developed CRISPR technology can help address these issues, having a power to perform treatment targeted to specific disease mechanisms with little to no adverse effects to the healthy tissue.Many gene therapies require intracellular delivery, and their efficacy can be highly influenced by the choice of the delivery vehicle. Due to their near-infrared fluorescence capabilities and large payload carrying capacity, carbon nanomaterials can be appropriate candidates for this role. Our work utilizes graphene quantum dots (GQDs) and single-walled carbon nanotubes (SWCNTs) as delivery vehicles for several CRISPR-Cas9-based formulations specifically designed to edit out genes related to immortalization and uncontrolled growth of cancer cells. Both of these nanomaterials exhibit visible and near-infrared fluorescence capabilities sufficient for in vitro and in vivo imaging, and, when bound to CRISPR-Cas9 system, are not only expected to transport the therapeutic, but also trace its delivery pathways. In order to warrant more efficient complexation with CRISPR-Cas9 system, GQDs are synthesized bottom-up from cationic polymers and characterized to exhibit stable fluorescence and high biocompatibility at ~1 mg/mL in vitro. Payload attachment is assessed microscopically and reflected by Zeta potential variation upon complexation. Both resulting nanomaterials-delivered formulations show successful target gene scission and intracellular tracking of the gene therapeutic via intrinsic nanomaterial fluorescence. The end goal of this work is to edit out and/or correct aberrant genes in cancer cells making them more susceptible to conventional treatment pathways, thus, ultimately helping improve cancer survival rates.