UC Santa Barbara

Intracellular delivery of biomolecules is hindered by both the size and charge of the biomolecule. While a number of delivery methods are available for genetic material, few are available for larger biomolecules like peptides and proteins. The delivery of these biomolecules are often limited to viral transfections of the genetic material which can lead to off-target gene editing due to prolonged expression of a protein within a cell. We developed and studied a delivery method for poly-histidine tagged proteins and peptides of variable sizes and charges for transient protein expression with light control via hollow gold nanoshells (HGNs). The construct of our delivery system involves a thiolated strand (nucleic acid or PEG based) containing an NTA attached to the particle’s gold surface. A his-tagged protein or peptide of interest is attached to the HGN via copper-NTA affinity. Endosomal uptake of these particles is mediated through an orthogonal strand containing a cell penetrating peptide (CPP). Protein release and endosomal disruption is achieved after irradiation with a focused femtosecond pulsed-laser of biologically benign near-infrared light. This thesis evaluates a variety of areas, from increasing the therapeutic efficacy of an apoptotic peptide to visualization of protein movement using live cell imaging with VIPERnano.