Methods & Techniques

While standard biochemistry and molecular biological techniques suffice for much of our work, several of our studies require sophisticated state-of-the-art equipment that is made available to us at the California NanoSystems Institute (CNSI) and the Molecular Biology Institute (MBI) here at UCLA or in the laboratories of our collaborators.

Physical/Chemical

Fluorescence correlation spectroscopy (FCS); high resolution cryo-electron microscopy (cryo-EM), including single-particle tomography; dynamic light scattering (DLS); magnetic and optical tweezers (Professors David Bensimon [ENS, Paris, and UCLA] and Doug Smith [UCSD]); maleimide , NHS ester, and “click” chemistries for conjugating ligands to capsid protein; electrophoretic, sedimentation, chromatographic and immunoblot separations and analyses of fluorescently-labeled RNA, protein, and RNA-protein complexes; labeling of RNA ends, and of capsid proteins, by small (typically, <3nm gold particles; liposome formulations for syntheses of enveloped virus-like particles; statistical-mechanical modeling.

Biological

Agrobacterial transformation of plants for high-level expression of wildtype and mutant CCMV capsid protein; bacterial expression systems for syntheses of capsid protein/targeting ligand fusions; genetic engineering of RNA-virus-derived replicons from mammalian, insect, and plant viruses; quantitative reverse-transcription PCR; transfection, infection and activation of cultured epithelial and dendritic cells for assays of RNA uptake, replication and protein expression levels; in vitro transcription and genetic engineering of viral genes and their protein products; cell-free translation of RNA, of virus-like particles, and of viruses.