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Principal Investigator: Gaurav Sahay
Organization: OREGON STATE UNIVERSITY
Fiscal Year: 2022
Award: $648,444
Funding agency: National Eye Institute
ABSTRACT
Inherited retinal dystrophies (IRDs) are a heterogenous group of orphan diseases, inherited in an autosomal
dominant, recessive or X-linked pattern in addition to mitochondrial transmission, all leading to the loss of
functional vision and often progressing to blindness. As a group, IRDs are due to mutations in over 280 genes.
Currently, there is only one FDA approved gene therapy for this large family of retinal degenerations. Prime
editing, a new versatile genome editing tool, allows for all 12 base-to-base changes, insertions up to 44 bases
long and deletions of up to 80 bases. Prime editors are capable of correcting 89% of known genetic variants
associated with human disease, but are still in their infancy for in-vivo use. Our long-term goal is to optimize
prime editing platforms for IRDs. Lipid based nanoparticles (LNPs) are a modular platform that can
encapsulate and deliver genome editors. Delivering nucleases as mRNA has been an optimal alternative
strategy for transient protein expression rather than persistent expression of DNA cutting machinery associated
with viral vectors. LNPs are capable of rapid and efficient delivery of mRNA to the retinal pigment epithelium,
however, they have limited capacity to transfect photoreceptors, which is necessary to target the many genes
associated with IRDs. We hypothesize that by employing phage display techniques, we will isolate promising
targeting peptides which will decorate our LNPs and effectively deliver prime editing cargo to the
photoreceptors. Our main goal is to generate peptide-targeted LNPs that lead to cell-specific delivery of prime
editing components for the treatment of IRDs. To achieve this goal, we propose the following specific aims: 1)
Optimize in-vivo phage display biopanning for the identification of targeting peptide moieties that allow for
photoreceptor-specific lipid nanoparticle-based gene delivery, 2) Dissect the mechanism of peptide-targeted
LNP entry into photoreceptors, and 3) Evaluate the efficacy, and any associated toxicity, of prime editing in two
rodent models of IRD. Thus far, we have identified novel peptides that can steer LNPs toward photoreceptor
gene delivery and determined that LNPs can package all prime editing components together and lead to
efficient prime editing of reporter genes in-vitro and in-vivo. Successful completion of this project will lead to the
development of cell-specific gene editing platforms that will advance treatment for IRDs.
Terms: <Adeno-Associated Viruses><Affect><Age><Amyloidosis><Assay><Bacteriophages><Bioassay><Biologic Assays><Biological><Biological Assay><Blindness><COVID-19><COVID19><CV-19><CV19><Cell Body><Cell Line><CellLine><Cells><DNA><DNA Nucleases><DNA Therapy><DNA delivery><DNase><Data><Deoxyribonucleases><Deoxyribonucleic Acid><Dependoparvovirus><Dependovirus><Development><Docking><Electroretinography><Encapsulated><FDA approved><Family><Fluorescence Activated Cell Sorting Fractionation><Fluorescence-Activated Cell Sorting><Fluorescence-Activated Cell Sortings><Frequencies><Gene Delivery><Gene Transfer Clinical><Gene variant><Genes><Genetic><Genetic Alteration><Genetic Change><Genetic Intervention><Genetic defect><Genome><Goals><Green Fluorescent Proteins><Hereditary><Immunoblotting><In Vitro><Inherited><Lead><Leucine Zippers><Libraries><Ligands><Link><Measurement><Measures><Mediating><Messenger RNA><Mice><Mice Mammals><Mitochondria><Modeling><Mouse Protein><Murine><Mus><Mutation><Neural Retina><Orphan Disease><Outer pigmented layer of retina><Patients><Pattern><Pb element><Peptide Receptor><Peptides><Persons><Phage Display><Phages><Photoreceptor Cell><Photoreceptors><Photosensitive Cell><Pigment cell layer of retina><Pigmented layer of retina><Population><Post-Transcriptional Gene Silencing><Posttranscriptional Gene Silencing><Prealbumin><Proalbumin><Proteins><Publishing><Quelling><RNA Interference><RNA Silencing><RNAi><Rare Diseases><Rare Disorder><Receptor Protein><Reporter Genes><Retina><Retina Proper><Retinal Degeneration><Retinal Dystrophy><Retinal Pigment Epithelium><Retinal pigment epithelial cells><Rodent Model><Sequence-Specific Posttranscriptional Gene Silencing><Sight><Strains Cell Lines><Structure of retinal pigment epithelium><Surface><System><Techniques><Thick><Thickness><Toxic effect><Toxicities><Transfection><Transmission><Transthyretin><Viral Vector><Vision><Visual Receptor><Western Blotting><Western Immunoblotting><adeno associated virus group><ages><allele variant><allelic variant><amyloid disease><bacterial virus><base><biologic><corona virus disease 2019><coronavirus disease 2019><coronavirus disease-19><coronavirus infectious disease-19><cultured cell line><deep sequencing><degenerative retina diseases><deliver DNA><developmental><efficacy analysis><efficacy assessment><efficacy evaluation><efficacy examination><electroretinogram><evaluate efficacy><examine efficacy><experiment><experimental research><experimental study><functional loss><gene repair therapy><gene therapy><gene-based therapy><genetic therapy><genetic variant><genome editing><genome mutation><genomic editing><genomic therapy><genomic variant><heavy metal Pb><heavy metal lead><human disease><in vivo><infancy><infantile><inherited retinal degeneration><knock-down><knockdown><lipid based nanoparticle><lipid nanoparticle><mRNA><mRNA delivery><mitochondrial><mouse model><murine model><nano particle delivery><nanoparticle delivered><nanoparticle delivery><novel><nuclease><orphan disorder><photoreceptor degeneration><prime editing><prime editor><protein blotting><protein expression><receptor><receptor mediated endocytosis><retina degeneration><retinal degenerative><retinal degenerative diseases><screening><sex><subretinal injection><tool><transmission process><uptake><vision loss><visual function><visual loss>