Structural Mechanisms Of Genome Flow In Bacteriophage T4 And Their Biomedical Applications
Structural Mechanisms Of Genome Flow In Bacteriophage T4 And Their Biomedical Applications
批准号:
10635661
负责人:
Venigalla B. Rao
金额:
$48.48万
依托单位国家:
美国
项目类别:
财政年份:
2023
资助国家:
美国
项目状态:
未结题
起止时间:
2023-02-15 至 2028-01-31
关键词:
Archaeal VirusesBacteriophage T4BacteriophagesBindingBiochemistryCapsidCategoriesCellsCessation of lifeComplexCryoelectron MicroscopyDNADNA PackagingDNA StructureDataData CollectionDefectDegenerative DisorderDiseaseDockingDuchenne muscular dystrophyDystrophinExperimental DesignsFutureGene DeliveryGenesGeneticGenomeHeadHumanIn VitroIndividualInfectionInterdisciplinary StudyInvestigationKnowledgeLengthLentivirusLife Cycle StagesLigandsMeasuresModelingMolecularMolecular ConformationMotorMuscleMuscle CellsMuscular DystrophiesMyopathyNanovirusNeckOrganismPhasePhysical condensationPlanet EarthPositioning AttributePreparationProcessProteinsRegulationReporter GenesResearchResearch ProposalsResolutionSeriesStructureSurfaceSystemTailTherapeuticTimeTranslatingTranslational ResearchTranslationsTubeViralViral GenomeVirusconformational conversiondelivery vehicledensitydesignds-DNAeffective therapyexperimental studyextracellulargene therapygenetic payloadhuman diseaseimprovedin vivoinnovationmouse modelmuscular dystrophy mouse modelmutantnanocapsidnanomachinenanoparticlenanoparticle deliverynovelnucleic acid binding proteinpressureprotein complexreconstructionsealtargeted deliverytherapeutic genevectorvirology
中文摘要
该建议旨在填补二十面体病毒组装的关键知识空白;机制和
英文摘要
This proposal aims to fill a critical knowledge gap in the assembly of icosahedral viruses; mechanisms and
controls by which a viral genome flows into and out of a virus capsid with high precision and fidelity. The tailed
dsDNA bacteriophage T4 is our model virus. The proposal will also translate this basic knowledge into a gene
therapy for muscular dystrophy, a debilitating degenerative muscular disease that causes early death.
We propose an innovative experimental design by advancing a novel “molecular valve” hypothesis. The
hypothesis states that a sophisticated molecular valve at the unique portal vertex controls genome flow into
and out of a virus capsid through dynamic structural and conformational changes.
By integrating genetics, biochemistry, and cryo-electron microscopy, we will generate a series of
asymmetric reconstructions of nanomachines in different structural and conformational states. These machines
translocate genome into virus capsid creating a pressurized condensate (inward genome flow), arrest genome
flow and position for delivery, and allow genome ejection upon encountering new host (outward genome flow).
In specific aim 1, we will generate atomic level structures of the DNA packaging machine consisting of
the portal vertex-bound packaging motor and its intermediate states during active translocation. A detailed
packaging mechanism will be formulated that will have broad implications to phages, eukaryotic and archaeal
viruses including herpes and adeno viruses. A strong work-flow has been established for preparation and data
collection of packaging complexes using a newly constructed super-charged “acidic capsid” mutant. In specific
aim 2, we will elucidate the dynamic mechanism of the neck-connector valve complex bound to portal by
generating structures in different assembly states. These structures would illustrate conformational changes in
the valve complex that lead to arrest of genome flow post-packaging and position it for delivery following tail
docking. We present a novel discovery involving the participation of a host nucleic acid binding protein Hfq in
these dynamic transactions. In specific aim 3, we will probe the mechanism of genome ejection by asymmetric
cryo-EM reconstructions of the genome ejection machine pre-infection, during-infection, and post-infection. A
preliminary cryo-EM reconstruction revealed, for the first time, density for a helical tape measure protein-DNA
complex in the innermost core of the ejection tube. In specific aim 4, we will incorporate the basic knowledge
gained from specific aims 1-3 to establish a novel, large capacity, T4 gene therapy vehicle to deliver the full-
length ~11-kb dystrophin gene into human muscle cells as well as into a muscular dystrophy mouse model.
Relying on our 42-years of expertise on T4 phage assembly and genome packaging, our synergistic
research team will uncover the basic mechanisms of genome flow in viruses and their translation into potentially
transformative phage-based gene therapeutics. These will have broad implications to virology and human
disease therapies.
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会议论文
Single Dose, Multivalent, Anthrax Plague Vaccines using Bacteriophage T4 Nanopart
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批准号:8819513
-
项目类别:
-
资助金额:$59.35万
-
财政年份:2014
-
负责人:Venigalla B. Rao
-
依托单位:
Single Dose, Multivalent, Anthrax Plague Vaccines using Bacteriophage T4 Nanopart
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批准号:8694624
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项目类别:
-
资助金额:$69.31万
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财政年份:2014
-
负责人:Venigalla B. Rao
-
依托单位:
Single Dose, Multivalent, Anthrax Plague Vaccines using Bacteriophage T4 Nanopart
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批准号:9000614
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项目类别:
-
资助金额:$63.63万
-
财政年份:2014
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负责人:Venigalla B. Rao
-
依托单位:
Potent Phage T4 Derived V2 Immunogens as HIV Vaccines
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批准号:8494569
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项目类别:
-
资助金额:$36.65万
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财政年份:2012
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负责人:Venigalla B. Rao
-
依托单位:
Potent Phage T4 Derived V2 Immunogens as HIV Vaccines
-
批准号:8685883
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项目类别:
-
资助金额:$48.54万
-
财政年份:2012
-
负责人:Venigalla B. Rao
-
依托单位:
Potent Phage T4 Derived V2 Immunogens as HIV Vaccines
-
批准号:8868023
-
项目类别:
-
资助金额:$47.13万
-
财政年份:2012
-
负责人:Venigalla B. Rao
-
依托单位:
Potent Phage T4 Derived V2 Immunogens as HIV Vaccines
-
批准号:8410257
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项目类别:
-
资助金额:$41.38万
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财政年份:2012
-
负责人:Venigalla B. Rao
-
依托单位:
Multivalent Plague, Anthrax Vaccines Using Bacteriophage T4 Display
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批准号:8435493
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项目类别:
-
资助金额:$72.6万
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财政年份:2009
-
负责人:Venigalla B. Rao
-
依托单位:
Multivalent Plague, Anthrax Vaccines Using Bacteriophage T4 Display
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批准号:7644596
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项目类别:
-
资助金额:$24.88万
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财政年份:2009
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负责人:Venigalla B. Rao
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依托单位:
Engineering a packaging nanomotor for delivery of RNA and other molecules
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批准号:7904763
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项目类别:
-
资助金额:$16.18万
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财政年份:2009
-
负责人:Venigalla B. Rao
-
依托单位:
Multivalent Plague, Anthrax Vaccines Using Bacteriophage T4 Display
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批准号:8232948
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项目类别:
-
资助金额:$53.03万
-
财政年份:2009
-
负责人:Venigalla B. Rao
-
依托单位:
Multivalent Plague, Anthrax Vaccines Using Bacteriophage T4 Display
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批准号:8036080
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项目类别:
-
资助金额:$47.45万
-
财政年份:2009
-
负责人:Venigalla B. Rao
-
依托单位:
Engineering a packaging nanomotor for delivery of RNA and other molecules
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批准号:7712849
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项目类别:
-
资助金额:$18.66万
-
财政年份:2009
-
负责人:Venigalla B. Rao
-
依托单位:
Multivalent Plague, Anthrax Vaccines Using Bacteriophage T4 Display
-
批准号:7783779
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项目类别:
-
资助金额:$25.2万
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财政年份:2009
-
负责人:Venigalla B. Rao
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依托单位:
A Multicomponent Anthrax Vaccine using Phage T4 Display
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批准号:7179265
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项目类别:
-
资助金额:$80.04万
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财政年份:2003
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负责人:Venigalla B. Rao
-
依托单位:
A Multicomponent Anthrax Vaccine using Phage T4 Display
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批准号:6792177
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项目类别:
-
资助金额:$66.67万
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财政年份:2003
-
负责人:Venigalla B. Rao
-
依托单位:
A Multicomponent Anthrax Vaccine using Phage T4 Display
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批准号:6689340
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项目类别:
-
资助金额:$31.68万
-
财政年份:2003
-
负责人:Venigalla B. Rao
-
依托单位:
A Multicomponent Anthrax Vaccine using Phage T4 Display
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批准号:6853629
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项目类别:
-
资助金额:$69.5万
-
财政年份:2003
-
负责人:Venigalla B. Rao
-
依托单位:
A Multicomponent Anthrax Vaccine using Phage T4 Display
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批准号:7012679
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项目类别:
-
资助金额:$82.58万
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财政年份:2003
-
负责人:Venigalla B. Rao
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依托单位:
STRUCTURE OF LARGE TERMINASE SUBUNIT FROM PHAGE T4
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批准号:6444683
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项目类别:
-
资助金额:$29.31万
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财政年份:2001
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负责人:Venigalla B. Rao
-
依托单位:
海外基金