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Principal Investigator: DANIEL G ANDERSON
Organization: MASSACHUSETTS INSTITUTE OF TECHNOLOGY
Fiscal Year: 2023
Award: $387,750
Funding agency: National Heart Lung and Blood Institute
Gene editing is a promising strategy for treating or even permanently curing genetic diseases. In
particular, a new technique called prime editing has the potential to make small targeted
insertions, deletions, and substitutions with very high potential coverage of known disease-
causing mutations, and while minimizing dangerous double-stranded breaks in DNA. In order to
realize this potential, robust delivery strategies must be developed to deliver prime editing tools
efficiently to disease-relevant organs. One such delivery strategy is lipid nanoparticle delivery of
RNA and/or protein-based prime editing components. LNPs are nonviral, nontoxic, and clinically
validated delivery tools. However, there is an extremely diverse space of possible LNPs, with
tens of thousands of potential lipid structures that may be useful for LNP delivery. Selecting the
best possible LNP for a prime editing application, therefore, is challenging because in vitro
testing is often unreliable and in vivo testing of one LNP at a time is extremely low throughput.
Here, we propose to combine two scalable techniques to generate and test safe, potent LNP
formulations for performing prime editing. First, we will employ combinatorial chemistry
techniques to generate large libraries of biodegradable lipids for inclusion into LNPs. Second,
we will introduce a new technique which we term pegRNA barcoding to screen dozens to
hundreds of LNPs for successful prime editing in a single mouse. We will employ this technique
to identify the best biodegradable LNPs for editing of multiple organs, including in particular the
lung and the liver. Having identified the top candidates, we will proceed to use our LNPs to
apply prime editing to treat mouse models of two different inherited genetic diseases: hereditary
tyrosinemia type I (HTI), a liver disease, and cystic fibrosis (CF), primarily a lung disease. We
will evaluate the efficiency of prime editing, the levels of undesired editing events, and
phenotypic correction of these mice. The results may identify promising preclinical candidates
for the treatment of HTI, CF, and many other lung and liver diseases.
Terms: <Active Follow-up><Bar Codes><Body Tissues><CFTR Mouse><CRISPR><CRISPR/Cas system><Cell Body><Cells><Chimera Protein><Chimeric Proteins><Clinical><Clinical Trials><Clustered Regularly Interspaced Short Palindromic Repeats><Combinatorial Chemistry Technics><Combinatorial Chemistry Techniques><Cystic Fibrosis><Cystic Fibrosis Transmembrane Conductance Regulator mouse><DNA><DNA Double Strand Break><DNA Nicking Enzyme><Dangerousness><Deoxyribonucleic Acid><Development><Disease><Disease model><Disorder><Drug Kinetics><EC 2.7.7.49><Encapsulated><Endonuclease I><Epithelium><Event><Fat Droplet><Formulation><Fumarylacetoacetase><Fusion Protein><Gene Delivery><Genes><Genetic Alteration><Genetic Change><Genetic Diseases><Genetic defect><Genome><Health><Hepatic Disorder><Hereditary Disease><Hereditary Tyrosinemias><Human><Human Genome><Hydrolase><Hydrolase Family Gene><Hydrolase Gene><In Vitro><Inborn Genetic Diseases><Inbred CFTR Mice><Individual><Inherited disorder><Intravenous><Libraries><Lipid Inclusion><Lipids><Liver><Liver diseases><Lung><Lung Respiratory System><Lung diseases><Measures><Mice><Mice Mammals><Mission><Modern Man><Mucoviscidosis><Murine><Mus><Mutation><NIH><National Institutes of Health><Nebulizer><Nickase><Non-Polyadenylated RNA><Organ><Pharmacokinetics><Phenotype><Progenitor Cells><Property><Proteins><Pulmonary Diseases><Pulmonary Disorder><RNA><RNA Gene Products><RNA Transcriptase><RNA delivery><RNA-Dependent DNA Polymerase><RNA-Directed DNA Polymerase><Reporting><Research><Reverse Transcriptase><Revertase><Ribonucleic Acid><Somatic Cell><Sorting><Structure><System><Techniques><Technology><Testing><Time><Tissues><Toxic effect><Toxicities><Tyrosinemias><United States National Institutes of Health><Writing><active followup><barcode><base editing><combinatorial chemistry><design><designing><developmental><disease of the lung><disease-causing mutation><disorder model><disorder of the lung><effective therapy><effective treatment><experiment><experimental research><experimental study><experiments><follow up><follow-up><followed up><followup><fumarylacetoacetate fumarylhydrolase><fumarylacetoacetate hydrolase><genetic condition><genetic disorder><genome editing><genome mutation><genomic editing><hepatic body system><hepatic disease><hepatic organ system><hepatopathy><hereditary disorder><heritable disorder><human whole genome><improved><in vitro testing><in vivo><in vivo evaluation><in vivo testing><inborn error><inherited diseases><inherited genetic disease><inherited genetic disorder><lipid based nanoparticle><lipid nanoparticle><liver disorder><lung disorder><mouse model><murine model><nano formulation><nano particle delivery><nanoformulation><nanoparticle delivered><nanoparticle delivery><nebulization><nebulize><new approaches><novel><novel approaches><novel strategies><novel strategy><pre-clinical><preclinical><prime editing><prime editor><pulmonary><stem cells><tool>