Document text
Principal Investigator: Mark Bathe
Organization: MASSACHUSETTS INSTITUTE OF TECHNOLOGY
Fiscal Year: 2019
Award: $193,750
Funding agency: National Institute of Biomedical Imaging and Bioengineering
PROJECT SUMMARY
Targeted delivery of therapeutic gene editing CRISPR ribonucleoprotein (RNP) complexes to cells offers major
opportunities for next-generation therapies, however, realization of this goal requires a vehicle to protect and
direct CRISPR RNPs to their targeted cells and tissues. Currently, there are no vehicles that can incorporate
CRISP RNPs for cellular delivery in vivo. Here, we propose to use structured DNA nanoparticles (DNA-NPs) as
versatile carriers for the targeted delivery of RNPs, taking advantage of base-pairing between the RNP and the
DNA-NP to facilitate RNP incorporation and controlled release. DNA-NPs will be designed with stoichiometric
control over the number of RNP payloads they carry internally, and functionalized externally with antibodies
and lipid binding moieties to control cellular targeting and intracellular trafficking. We will minimize macrophage
engulfment and enhance blood compatibility through surface functionalization with polyethylene glycol (PEG)
and other biocompatible passivation techniques, using standard assays for nanoparticle physiological and
immunological interactions. Binding and release of CRISPR RNPs from DNA-NPs will be assayed in vitro, and
DNA-RNP base pairing will ensure stable binding and facile release of cargo from vehicle. Genome editing will
be assayed through both next generation sequencing and targeted PCR, which will esure that the enzymatic
activity of the cargo remains intact during delivery. Targeting CRISPR RNPs to sub-populations of cultured
cells expressing cell specific surface receptors will be evaluated, with particular attention paid targeted gene
editing in cells of interest. The generality of our RNP delivery platform offers a unique and transformative
approach to treating a range of deadly and currently untreatable genetic diseases and cancers by targeted
delivery of CRISPR.
Terms: <3-D structure><3-dimensional structure><3D structure><Antibodies><Assay><Attention><Base Pairing><Binding><Bioassay><Biocompatible Materials><Biologic Assays><Biological Assay><Biological Markers><Biomaterials><Blood><Blood Cells><Blood Plasma><Blood Reticuloendothelial System><Body Tissues><CRISPR><CRISPR/Cas system><Cancers><Cell Body><Cells><Chemical Structure><Chemicals><Chemotactic Cytokines><Clotting><Clustered Regularly Interspaced Short Palindromic Repeats><Coagulation><Coagulation Process><Complement Activation><Complex><Cultured Cells><DNA><DNA Structure><DNA Therapy><Deoxyribonucleic Acid><Development><Disease><Disorder><Ensure><Enzyme Gene><Enzymes><Future><Gene Delivery><Gene Transfer Clinical><Gene-Modified><Genes><Genetic Diseases><Genetic Intervention><Genomic DNA><Geometry><Goals><Health><Hemolysis><Hereditary><Homologous Chemotactic Cytokines><Human><IFN><Immunochemical Immunologic><Immunologic><Immunological><Immunologically><Immunologics><Immunology procedure><In Vitro><Inherited><Innate Immune Response><Intercrines><Interferons><Ligands><Lipid Binding><Macrogols><Malignant Neoplasms><Malignant Tumor><Messenger RNA><Modern Man><Modification><Molecular Interaction><Mononuclear><NGS Method><NGS system><Nucleic Acids><Peripheral Blood Cell><Phagocytes><Phagocytic Cell><Physiologic><Physiological><Plasma><Plasma Serum><Platelet aggregation><Polyethylene Glycols><Polyethylene Oxide><Polyethyleneoxide><Polymers><Polyoxyethylenes><Receptor Protein><Reticuloendothelial System, Serum, Plasma><Ribonucleoproteins><SIS cytokines><Short interfering RNA><Small Interfering RNA><Structure-Activity Relationship><Surface><Survey Instrument><Surveys><System><Techniques><Technology><Testing><Therapeutic><Therapeutic Uses><Time><Tissues><Toxic effect><Toxicities><Viral><amebocyte><bio-markers><biocompatibility><biologic marker><biological material><biomarker><biomaterial compatibility><cellular targeting><chemical property><chemical structure function><chemoattractant cytokine><chemokine><complement pathway regulation><controlled release><cytokine><design><designing><developmental><erythrolysis><experiment><experimental research><experimental study><gDNA><gene modification><gene therapeutics><gene therapy><gene-based therapeutics><gene-based therapy><genes therapeutic><genes therapeutics><genetic condition><genetic disorder><genetic therapy><genome editing><genome sequencing><genomic editing><genomic therapy><immunogenic><immunologic assay><immunologic assay/test><in vitro Assay><in vivo><interest><lipid bound><mRNA><macrophage><malignancy><nano particle><nano particle delivery><nano-sized particle><nanoparticle><nanoparticle delivered><nanoparticle delivery><nanosized particle><neoplasm/cancer><next gen sequencing><next generation><next generation sequencing><nextgen sequencing><physical property><programs><prophylactic><receptor><response><siRNA><site targeted delivery><structure function relationship><targeted delivery><therapeutic RNA><therapeutic gene><therapeutic nanoparticles><three dimensional structure><tool><trafficking><uptake>