grant

Extracellular vesicles encapsulating CRISPR machinery for treatment of SARS-CoV-2 infection

Organization UNIVERSITY OF GEORGIALocation ATHENS, UNITED STATESPosted 19 Jan 2023Deadline 31 Dec 2026
NIHUS FederalResearch GrantFY20242019 novel corona virus2019 novel coronavirus2019-nCoV2019-nCoV S protein2019-nCoV spike glycoprotein2019-nCoV spike proteinACE2Aminoacetic AcidAnti-viral AgentsBCAR1BCAR1 ProteinBCAR1 geneBindingBiologicalBiologyBody TissuesBreast Cancer Anti-Estrogen Resistance 1 ProteinCKRAS proteinCOVID infected patientCOVID patientCOVID positive patientCOVID-19COVID-19 S proteinCOVID-19 infected patientCOVID-19 infectionCOVID-19 patientCOVID-19 positive patientCOVID-19 spikeCOVID-19 spike glycoproteinCOVID-19 spike proteinCOVID-19 therapyCOVID-19 treatmentCOVID-19 virusCOVID-19 virus infectionCOVID19 infectionCOVID19 patientCOVID19 positive patientCOVID19 virusCRISPRCRISPR approachCRISPR based approachCRISPR methodCRISPR methodologyCRISPR techniqueCRISPR technologyCRISPR toolsCRISPR-CAS-9CRISPR-based methodCRISPR-based techniqueCRISPR-based technologyCRISPR-based toolCRISPR/CAS approachCRISPR/Cas methodCRISPR/Cas systemCRISPR/Cas technologyCRISPR/Cas9CRISPR/Cas9 technologyCRK-Associated SubstrateCRKASCV-19CarbonCas nuclease technologyCas proteinCause of DeathCell BodyCell CommunicationCell InteractionCell membraneCell-to-Cell InteractionCellsClustered Regularly Interspaced Short Palindromic RepeatsClustered Regularly Interspaced Short Palindromic Repeats approachClustered Regularly Interspaced Short Palindromic Repeats methodClustered Regularly Interspaced Short Palindromic Repeats methodologyClustered Regularly Interspaced Short Palindromic Repeats techniqueClustered Regularly Interspaced Short Palindromic Repeats technologyCoV-2CoV2Coated vesicleComplexConditioned Culture MediaConditioned MediumCoronavirus Infectious Disease 2019Cytoplasmic MembraneDataEncapsulatedEngineeringEpithelial CellsEventFutureGene ExpressionGene ModifiedGenerationsGenesGeneticGenetic DiseasesGenetic EngineeringGenetic Engineering BiotechnologyGenetic Engineering Molecular BiologyGlycineGolden HamstersGolden Syrian HamstersIn VitroKnock-outKnockoutKnowledgeLipidsLungLung ParenchymaLung Respiratory SystemLung TissueMediatingMembraneMesocricetus auratusMessenger RNAMethodsMicro RNAMicroRNAsModelingModificationMolecularMolecular InteractionN-terminalNH2-terminalNon-Polyadenylated RNANucleocapsidPersonsPlasma MembranePlayProductionProliferatingProteinsRNARNA Gene ProductsRNA SequencesReceptor ProteinRecombinant DNA TechnologyRespiratory EpitheliumRibonucleic AcidRibonucleoproteinsRoleSARS corona virus 2SARS-CO-V2SARS-COVID-2SARS-CoV-2SARS-CoV-2 SSARS-CoV-2 S proteinSARS-CoV-2 infected patientSARS-CoV-2 infectionSARS-CoV-2 inhibitorSARS-CoV-2 patientSARS-CoV-2 positive patientSARS-CoV-2 spikeSARS-CoV-2 spike glycoproteinSARS-CoV-2 spike proteinSARS-CoV-2 therapySARS-CoV-2 treatmentSARS-CoV2SARS-CoV2 infectionSARS-associated corona virus 2SARS-associated coronavirus 2SARS-coronavirus-2SARS-related corona virus 2SARS-related coronavirus 2SARSCoV2Saturated Fatty AcidsSevere Acute Respiratory Coronavirus 2Severe Acute Respiratory Distress Syndrome CoV 2Severe Acute Respiratory Distress Syndrome Corona Virus 2Severe Acute Respiratory Distress Syndrome Coronavirus 2Severe Acute Respiratory Syndrome CoV 2Severe Acute Respiratory Syndrome-associated coronavirus 2Severe Acute Respiratory Syndrome-related coronavirus 2Severe acute respiratory syndrome associated corona virus 2Severe acute respiratory syndrome coronavirus 2Severe acute respiratory syndrome coronavirus 2 S proteinSevere acute respiratory syndrome coronavirus 2 infectionSevere acute respiratory syndrome coronavirus 2 inhibitorSevere acute respiratory syndrome coronavirus 2 spike glycoproteinSevere acute respiratory syndrome coronavirus 2 spike proteinSevere acute respiratory syndrome related corona virus 2Severity of illnessSpecificityStructure of parenchyma of lungStructure of respiratory epitheliumSyrian HamstersSystemTechnologyTestingTherapeutic Gene EditingTissuesTransfectionTransmissionVaccinesVariantVariationViralViral BurdenViral Gene ProductsViral Gene ProteinsViral LoadViral Load resultViral ProteinsVirus AssemblyWuhan coronavirusairway epitheliumangiotensin converting enzyme 2angiotensin converting enzyme IIanti-viral compoundanti-viral drugsanti-viral medicationanti-viral therapeuticanti-viralsbiologicblock SARS-CoV-2block severe acute respiratory syndrome coronavirus 2coronavirus disease 2019coronavirus disease 2019 S proteincoronavirus disease 2019 infected patientcoronavirus disease 2019 infectioncoronavirus disease 2019 patientcoronavirus disease 2019 positive patientcoronavirus disease 2019 spike glycoproteincoronavirus disease 2019 spike proteincoronavirus disease 2019 therapycoronavirus disease 2019 treatmentcoronavirus disease 2019 viruscoronavirus disease infected patientcoronavirus disease patientcoronavirus disease positive patientcoronavirus disease-19coronavirus disease-19 patientcoronavirus disease-19 viruscoronavirus infectious disease-19coronavirus patientdisease severityextracellular vesiclesflu infectionflu virus infectiongene modificationgene-editing therapygenetic conditiongenetic disordergenetically engineeredgenetically modifiedgenome editinggenome editing based therapygenome editing therapygenome editing treatmentgenome editing-based therapeuticsgenomic RNAgenomic editinghCoV19improvedin vivoinfected with COVID-19infected with COVID19infected with SARS-CoV-2infected with SARS-CoV2infected with coronavirus disease 2019infected with fluinfected with flu virusinfected with influenzainfected with influenza virusinfected with severe acute respiratory syndrome coronavirus 2influenza infectioninfluenza virus infectioninhibit SARS-CoV-2inhibit severe acute respiratory syndrome coronavirus 2mRNAmembrane structuremiRNAmiRNAsmortalitymyristoylationnCoV2nano particlenano-sized particlenanoparticlenanosized particlep130 cas proteinp130CASpandemicpandemic diseasepatient infected with COVIDpatient infected with COVID-19patient infected with SARS-CoV-2patient infected with coronavirus diseasepatient infected with coronavirus disease 2019patient infected with severe acute respiratory syndrome coronavirus 2patient with COVIDpatient with COVID-19patient with COVID19patient with SARS-CoV-2patient with coronavirus diseasepatient with coronavirus disease 2019patient with severe acute respiratory distress syndrome coronavirus 2plasmalemmaposttranscriptionalpulmonaryreceptorrespiratory tract epitheliumsevere acute respiratory syndrome coronavirus 2 infected patientsevere acute respiratory syndrome coronavirus 2 patientsevere acute respiratory syndrome coronavirus 2 positive patientsevere acute respiratory syndrome coronavirus 2 therapysevere acute respiratory syndrome coronavirus 2 treatmentsocial rolespike proteins on SARS-CoV-2src Kinasessrc Protein-Tyrosine Kinasessrc Tyrosine Kinasessrc-Family Kinasessrc-Family Tyrosine Kinasestherapeutic editingtherapeutic genome editingtransmission processtreat COVID-19treat SARS-CoV-2treat coronavirus disease 2019treat severe acute respiratory syndrome coronavirus 2treatment strategyviral assemblyviral genomicsvirus genomicsvirus protein
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Full Description

Summary/Abstract
SARS-CoV-2 has caused the deaths of millions of people globally. Effective antiviral therapeutic
treatment options are urgently needed. CRISPR-mediated genome editing has provided a very promising avenue
for treatment of a variety of genetic diseases. Particularly, the CRISPR-Cas13 system has been demonstrated
to possess the potential of inhibiting SARS-CoV-2 and influenza infections by degradation of viral genomic RNA
and viral mRNA. However, it is still very challenging to deliver the CRISPR machinery to initiate genome editing
efficiently in vivo. Extracellular vesicles (EVs) contain a variety of molecular components including lipids, mRNA,
microRNAs, and proteins. A large body of studies has shown that EVs mediate cell-to-cell communication by
transmitting their encapsulated contents. This proposal intends to construct EVs encapsulating the CRISPR
machinery and deliver the EVs to respiratory epithelial cells to inhibit SARS-CoV-2 proliferation in vivo. We will
therefore genetically engineer the Cas13d protein so that Cas13d/CRISPR-RNA (crRNA) ribonucleoprotein
complex can be encapsulated into EVs. We will also engineer the membrane of EVs, such that EVs target
respiratory epithelial cells and deliver Cas13d/crRNA for inhibiting SARS-CoV-2 viral assembly and proliferation,
thereby inhibiting COVID-19. This study will allow us to understand the feasibility of an EVs-based vehicle to
deliver genome editing machinery to inhibit SARS-CoV-2 proliferation in lung epithelial cells. This study will
provide a treatment option for COVID-19 patients to reduce disease severity and mortality.

Grant Number: 5R21AI171944-02
NIH Institute/Center: NIH
Principal Investigator: Houjian Cai

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