RNA nanoparticles and nanotubes
Inventors
Shapiro, Bruce A. • Yingling, Yaroslava G. • Bindewald, Eckart • Kasprzak, Wojciech • Jaeger, Luc • Severcan, Isil • Geary, Cody • Afonin, Kirill
Assignees
University of California San Diego UCSD • US Department of Health and Human Services
Publication Number
US-9732337-B2
Publication Date
2017-08-15
Expiration Date
2030-06-16
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Abstract
The instant invention provides polyvalent RNA nanoparticles comprising RNA motifs as building blocks that can form RNA nanotubes. The polyvalent RNA nanoparticles are suitable for therapeutic or diagnostic use in a number of diseases or disorders.
Core Innovation
The invention provides polyvalent RNA nanoparticles comprising RNA motifs as building blocks that can form RNA nanotubes. These RNA nanoparticles are capable of self-assembling into predefined sizes and geometrical shapes, including three-dimensional RNA polyhedral cages, which can carry multiple components such as molecules for specific cell recognition, image detection, and therapeutic treatment. The nanoparticles can encapsulate small therapeutic molecules inside their cages and release them upon triggering by small ligands.
The RNA motifs used as building blocks include diverse motifs such as RNA I or RNA II motifs, right angle (RA) motifs, three way junction (3WJ) motifs, four way junction motifs, and class II tRNA motifs. The nanoparticles can be spatially addressable by optimizing tail connectors and controlling the positioning of agents like biotin within the cage. These nanoparticles are designed to be multifunctional, capable of carrying therapeutic, diagnostic, and delivery agents, and can be manufactured both in vitro and in cells, assembling under conditions suitable for RNA.
The problem being solved addresses the limitations of current nanoparticle drug delivery systems, including poor stability, low assembly yields, undesired immune response, and difficulty in specific targeting of therapeutic agents. RNA nanoparticles offer advantages over DNA and protein nanoparticles, including decreased immune response, programmable self-assembly, versatility in function such as including aptamers and ribozymes, and better in vivo expression efficiency. The invention aims to provide a safe and efficient nanoparticle capable of delivering multiple therapeutic and diagnostic agents specifically to targeted cells or tissues.
Claims Coverage
The patent includes multiple independent claims focusing on synthetic polyvalent RNA nanocubes built from specific RNA motifs and methods for making and using these nanocubes. The inventive features cover the compositions, structural elements, conjugation with agents, and methods of synthesis and use.
Synthetic polyvalent RNA nanocube comprising specific 90 degree angle bend RNA motifs
A synthetic polyvalent RNA nanocube assembled using approximately 90 degree angle bend RNA motifs as building blocks, specifically selected from right angle (RA) motifs, three way junction (3WJ) motifs, four way junction motifs, and class II tRNA motifs, with RA motifs particularly comprising SEQ ID NO: 1-4.
Incorporation of defined 3WJ and class II tRNA motifs
The RNA nanocube can incorporate three way junction (3WJ) motifs selected from SEQ ID NO: 5-8 and class II tRNA motifs selected from SEQ ID NO: 9-16, allowing defined structural diversity in the nanocube assembly.
Nanocube comprising therapeutic, imaging, or diagnostic agent conjugation
The polyvalent RNA nanocube comprising one or more agents such as therapeutic, imaging, or diagnostic agents conjugated to the nanoparticle to provide multifunctionality.
Drug delivery composition using the polyvalent RNA nanocube
A drug delivery composition comprising the polyvalent RNA nanocube capable of gaining entry into cells or tissues, with optional inclusion of therapeutic agents or biotin for targeted delivery or functionalization.
Methods for making polyvalent RNA nanocubes
Methods involving either overexpressing RNA sequences in a cell comprising the defined 90 degree angle bend RNA motifs or mixing RNA sequences in vitro followed by heating and cooling steps to allow self-assembly into polyvalent RNA nanocubes.
Kit comprising polyvalent nanocubes and instructions
A kit containing the polyvalent RNA nanocube according to the invention along with instructions for use.
Nanocube structure comprising six or ten strands and specific structural features
The polyvalent RNA nanocube structure consisting of either six or ten RNA strands and optionally including at least one 5′ dangling end and connector sizes between seven to ten base pairs to define structural connectivity and stability.
The claims collectively cover synthetic polyvalent RNA nanocubes constructed from defined RNA motifs, their assembly into nanostructures with specific strand compositions, conjugation with multiple functional agents, drug delivery compositions using these nanocubes, and methods and kits for their production and use.
Stated Advantages
The RNA nanoparticles may not induce significant immune responses compared to protein nanoparticles.
They are smaller than many available nanoparticles, allowing increased efficiency of administration.
The nanoparticles can carry multiple and different therapeutic or diagnostic agents simultaneously.
RNA nanoparticles provide increased protection against ribonuclease degradation compared to monomeric RNAs.
The nanoparticles possess tunable thermodynamic and self-assembly properties based on the complexity of RNA motifs used.
RNA nanocubes have higher thermal stability compared to DNA counterparts.
The nanoparticles assemble isothermally under physiologic conditions, facilitating potential in vivo production.
Documented Applications
Therapeutic use in delivering multiple therapeutic and diagnostic agents specifically to target cells and tissues.
Drug delivery, including encapsulation and controlled release of therapeutic molecules.
Use as diagnostic tools or biosensors, including fluorescent molecular beacons and aptamer-based sensors.
Generation of nanoscale scaffolds for positioning proteins or functional RNAs for nanomedicine and synthetic biology.
Use in treating a variety of diseases such as cancer, AIDS, Alzheimer's, diabetes, asthma, arthritis, Parkinson's, and many others.
Functional nanostructures for imaging, gene therapy, cell targeting, and diagnostics.
Construction of complex supramolecular RNA assemblies (e.g., nanorings, nanotubes, nanogrids, cuboids, triads, nanoarrays).
Use as delivery vehicles that avoid immune clearance and allow repeated long-term treatment with low toxicity.
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