A Novel Hybrid Viral Vector for Therapeutic CRISPR Insertion of Multiple Transgenes

NIH Pandemic-Era Grants

Pandemic Era Grants

2024

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Principal Investigator: Logan Thrasher Collins
Organization: WASHINGTON UNIVERSITY
Fiscal Year: 2024
Award: $48,974
Funding agency: National Institute on Aging

PROJECT SUMMARY
Existing delivery systems for CRISPR-Cas gene editing machinery have very little capacity for making multigenic
insertions. As such, the scope of existing gene therapies has been limited primarily to monogenic diseases. Yet
multigenic insertions could facilitate treatment of a much broader range of conditions. To address this challenge,
I am developing a chimeric AdAAV gene editing delivery system by physically linking multiple adeno-associated
viruses (AAVs) to the surface of adenovirus (Ad). I am using the SpyTag-SpyCatcher technology for site-specific
covalent conjugation. In my design, the Ad will encode the Cas9 protein and the gRNAs while the AAVs will
provide a single-stranded DNA template. To enable multigenic genetic insertions, I will conjugate a mixture of
AAVs (with distinct genetic cargos) onto the Ads. Each AAV cargo sequence will undergo programmable
insertion into a unique genomic target site via the Cas9 and gRNAs encoded in the Ad. I anticipate that the strong
and targetable transduction of the Ad will efficiently drive the uptake of AdAAV particles into cells. I also anticipate
that greater in vivo gene editing efficiencies may occur using this design because of the known benefits of single-
stranded DNA templates for homology-directed repair (HDR). My fully novel AdAAV vector will act as an
efficacious and versatile platform for multigenic gene therapy. AdAAVs will be particularly suited for treating
aging and aging-related conditions, especially Alzheimer’s disease.

Terms: <AD dementia><Abbreviations><Address><Adeno-Associated Viruses><Adenoviridae><Adenoviruses><Aging><Alzheimer Type Dementia><Alzheimer disease dementia><Alzheimer sclerosis><Alzheimer syndrome><Alzheimer's><Alzheimer's Disease><Alzheimers Dementia><Amyloidosis><Avidity><BCAR1><BCAR1 Protein><BCAR1 gene><Blindness><Breast Cancer Anti-Estrogen Resistance 1 Protein><CKRAS protein><CRISPR><CRISPR based therapeutics><CRISPR based treatment><CRISPR therapeutics><CRISPR treatment><CRISPR-Cas based therapeutics><CRISPR-based disease therapeutics><CRISPR-based therapy><CRISPR/Cas system><CRISPR/Cas therapeutics><CRISPR/Cas9 therapeutics><CRISPR/Cas9 therapy><CRISPR/Cas9 treatment><CRISPR/Cas9-based therapy><CRK-Associated Substrate><CRKAS><Capsid><Cas protein><Cas9 based therapeutics><Cell Body><Cell surface><Cells><Clinical Trials><Clustered Regularly Interspaced Short Palindromic Repeats><Clustered Regularly Interspaced Short Palindromic Repeats based therapeutics><Clustered Regularly Interspaced Short Palindromic Repeats therapeutics><Complex><DNA><DNA Packaging><DNA Therapy><Data><Deoxyribonucleic Acid><Dependoparvovirus><Dependovirus><Diameter><Dose><Electron Microscopy><Fiber><Gene Delivery><Gene Proteins><Gene Therapy Vectors><Gene Transduction Agent><Gene Transduction Vectors><Gene Transfer Clinical><Genes><Genetic><Genetic Intervention><Genome><Genomics><Guide RNA><Hepatic Disorder><Hybrids><Leber congenital amaurosis><Leber's amaurosis><Leber's congenital amaurosis><Lentivirinae><Lentivirus><Link><Liver diseases><Location><Mendelian disease><Mendelian disorder><Mendelian genetic disorder><Position><Positioning Attribute><Prealbumin><Primary Senile Degenerative Dementia><Proalbumin><Protein Gene Products><Proteins><Protocol><Protocols documentation><Renaissance><Research><Risk><Side><Single-Stranded DNA><Site><Specific qualifier value><Specified><Structure><Surface><System><Technology><Therapeutic Effect><Therapeutic Gene Editing><Transgenes><Transthyretin><Viral Vector><adeno associated virus group><aging associated><aging related><amaurosis congenita of Leber><amyloid disease><cell transduction><cellular targeting><cellular transduction><congenital amaurosis of retinal origin><delivery vector><delivery vehicle><design><designing><experience><gRNA><gene repair therapy><gene therapy><gene-based therapy><gene-editing therapy><genetic therapy><genome editing based therapy><genome editing therapy><genome editing treatment><genome editing-based therapeutics><genomic therapy><hepatic disease><hepatopathy><in vivo><lipid based nanoparticle><lipid nanoparticle><liver disorder><microscope imaging><microscopic imaging><microscopy imaging><monogenic disease><monogenic disorder><new technology><novel><novel technologies><p130 cas protein><p130CAS><particle><pre-clinical><preclinical><prevent><preventing><primary degenerative dementia><prototype><repair><repaired><senile dementia of the Alzheimer type><single-gene disease><single-gene disorder><ssDNA><technology platform><technology system><therapeutic editing><therapeutic genome editing><transduced cells><transduction efficiency><transgene><uptake><vector><vision loss><visual loss>