Protein/oligonucleotide core-shell nanoparticle therapeutics
Inventors
Mirkin, Chad A. • Brodin, Jeffrey D.
Assignees
Publication Number
US-12264344-B2
Publication Date
2025-04-01
Expiration Date
2035-08-19
Interested in licensing this patent?
MTEC can help explore whether this patent might be available for licensing for your application.
Abstract
The present disclosure is directed to core-shell nanoparticles, compositions comprising core-shell nanoparticles, and methods of their use.
Core Innovation
The invention provides core-shell nanoparticles in which a single protein core is chemically functionalized with a dense shell of polynucleotides, such as DNA or RNA. This creates protein/oligonucleotide core-shell nanoparticles that can be used to construct crystalline materials from catalytically active proteins. The process uses covalent attachment of oligonucleotides to surface functional groups of proteins, forming a dense and highly anionic surface that can improve stability and cellular properties.
The dense oligonucleotide shell prevents aggregation and unfolding of the functionalized proteins and serves as a universal scaffold for further DNA-directed assembly. This programmable approach allows the construction of ordered supramolecular lattices using protein building blocks and enables predictable assembly of multi-component systems, including combinations of proteins and inorganic nanoparticles such as gold nanoparticles.
The problem addressed by the invention is the limited therapeutic potential of proteins due to poor cellular uptake, activation of the innate immune response, poor bioavailability, degradation by proteases, and aggregation or inactivation during storage. Existing DNA-mediated assembly strategies were primarily restricted to hard inorganic nanoparticles and did not offer the flexibility to exploit the functional diversity and monodispersity of proteins. The invention overcomes these limitations by providing a generalizable method of functionalizing proteins with oligonucleotides and assembling them into functional materials.
Claims Coverage
There are two independent method claims covering the inventive features regarding gene expression inhibition using protein/oligonucleotide core-shell nanoparticles.
Inhibiting gene expression using nanoparticle with protein core and oligonucleotide shell
A method comprising contacting a target polynucleotide with a nanoparticle that has: - a core consisting of a single protein - a shell consisting of a plurality of polynucleotides - a linker connecting the core to at least one polynucleotide in the shell, where the linker comprises Formula (I), (II), or both This method specifically results in inhibition of expression of a gene product encoded by the target polynucleotide.
Inhibiting gene expression using a nanoparticle having a linker of defined chemical structure
A method comprising contacting a target polynucleotide with a nanoparticle having: - a core comprised of a single protein - a shell of a plurality of polynucleotides - a linker between the core and at least one polynucleotide in the shell - the linker is selected from Formula (I), (II), or both specific chemical linkers as described This method achieves inhibition of the gene product encoded by the target polynucleotide due to the defined covalent attachment facilitated by the specified linker.
The claims cover methods for inhibiting gene expression using nanoparticles comprised of a single protein core and dense oligonucleotide shell, where covalent attachment is achieved via specified linkers, enabling targeted gene regulation.
Stated Advantages
The oligonucleotide shell imparts stability on the protein core, preventing aggregation and unfolding, and limits degradation by proteases.
Dense oligonucleotide functionalization promotes efficient cell entry and minimizes immune response compared to unmodified proteins.
The invention allows programmable assembly of proteins into ordered supramolecular materials with defined ratios and orientations.
The approach is generalizable to any protein, does not require genetic manipulation of the protein core, and supports assembly of lattices with multiple components.
The protein core provides native enzymatic or functional activity, while the oligonucleotide shell can contribute gene regulation or therapeutic responses.
The oligonucleotide sequence can be designed for controlled degradation rates, facilitating precise control over the exposure and function of the protein core in physiological settings.
The multiple functional groups on the protein surface allow orthogonal chemical modifications, enabling co-functionalization with additional targeting or therapeutic agents.
The oligonucleotide shell enhances cellular uptake and reduces immunogenic response compared to administration of protein alone.
Documented Applications
Stabilization of protein-based therapeutics against degradation by proteases or during prolonged storage.
Delivery of therapeutic proteins to cells, improving cellular uptake and reducing immunogenicity.
Replacement of deficient enzymes, such as in lysosomal storage disorders.
Enzymatic conversion of toxic metabolites, like catalase-mediated decomposition of hydrogen peroxide after traumatic injury or stroke.
General method for cellular delivery of oligonucleotides and proteins for multifunctional therapeutic strategies.
Assembly of proteins into single component or multi-component crystalline superlattices for catalysis or sensing.
Preparation of protein crystals suitable for X-ray structure determination.
Construction of tandem catalytic systems by combining multiple proteins in defined stoichiometry and orientation.
Use in detection of target molecules in vitro or in vivo, including gene expression assays or molecular diagnostics.
Gene regulation and inhibition of expression of target genes through hybridization of the oligonucleotide shell with target polynucleotides.
Combination of protein functionalization for cell targeting, e.g., inclusion of Mannose 6-phosphate for lysosomal targeting.
Interested in licensing this patent?