title: Tunable protease-activatable virus nanonodes.
abstract: We explored the unique signal integration properties of the self-assembling 60-mer protein capsid of adeno-associated virus (AAV), a clinically proven human gene therapy vector, by engineering proteolytic regulation of virus-receptor interactions such that processing of the capsid by proteases is required for infection. We find the transfer function of our engineered protease-activatable viruses (PAVs), relating the degree of proteolysis (input) to PAV activity (output), is highly nonlinear, likely due to increased polyvalency. By exploiting this dynamic polyvalency, in combination with the self-assembly properties of the virus capsid, we show that mosaic PAVs can be constructed that operate under a digital AND gate regime, where two different protease inputs are required for virus activation. These results show viruses can be engineered as signal-integrating nanoscale nodes whose functional properties are regulated by multiple proteolytic signals with easily tunable and predictable response surfaces, a promising development toward advanced control of gene delivery.
authors.affiliation: Department of Bioengineering and ‡Department of Biochemistry and Cell Biology, Rice University , Houston, Texas 77005, United States.
authors.forename: Justin
authors.initials: J
authors.lastname: Judd
authors.forename: Michelle L
authors.initials: ML
authors.lastname: Ho
authors.forename: Abhinav
authors.initials: A
authors.lastname: Tiwari
authors.forename: Eric J
authors.initials: EJ
authors.lastname: Gomez
authors.forename: Christopher
authors.initials: C
authors.lastname: Dempsey
authors.forename: Kim
authors.initials: K
authors.lastname: Van Vliet
authors.forename: Oleg A
authors.initials: OA
authors.lastname: Igoshin
authors.forename: Jonathan J
authors.initials: JJ
authors.lastname: Silberg
authors.forename: Mavis
authors.initials: M
authors.lastname: Agbandje-McKenna
authors.forename: Junghae
authors.initials: J
authors.lastname: Suh
mesh_terms: D000595:Amino Acid Sequence; D002213:Capsid / Q000737:chemistry; D000229:Dependovirus / Q000737:chemistry*; D018014:Gene Transfer Techniques; D015316:Genetic Therapy / Q000379:methods; D005822:Genetic Vectors; D049452:Green Fluorescent Proteins / Q000737:chemistry; D057809:HEK293 Cells; D006801:Humans; D008854:Microscopy, Electron; D008969:Molecular Sequence Data; D050997:Nanomedicine / Q000379:methods; D053758:Nanoparticles / Q000737:chemistry*; D036103:Nanotechnology / Q000379:methods*; D010447:Peptide Hydrolases / Q000737:chemistry*; D010957:Plasmids / Q000737:chemistry; D015202:Protein Engineering / Q000379:methods; D013499:Surface Properties; D013696:Temperature; D019076:Transgenes; D014780:Viruses / Q000737:chemistry*
chemical_list: D049452:Green Fluorescent Proteins; D010447:Peptide Hydrolases