Publication Date
6-11-2021
Journal
Molecular Therapy Methods & Clinical Development:
DOI
10.1016/j.omtm.2021.04.011
PMID
34141821
PMCID
PMC8166646
PubMedCentral® Posted Date
4-24-2021
PubMedCentral® Full Text Version
Post-print
Published Open-Access
yes
Keywords
AAV, CRISPR-Cas9, gene therapy, genome editing, gene targeting, Apoa1, liver, inherited metabolic disorders
Abstract
Clinical application of somatic genome editing requires therapeutics that are generalizable to a broad range of patients. Targeted insertion of promoterless transgenes can ensure that edits are permanent and broadly applicable while minimizing risks of off-target integration. In the liver, the Albumin (Alb) locus is currently the only well-characterized site for promoterless transgene insertion. Here, we target the Apoa1 locus with adeno-associated viral (AAV) delivery of CRISPR-Cas9 and achieve rates of 6% to 16% of targeted hepatocytes, with no evidence of toxicity. We further show that the endogenous Apoa1 promoter can drive robust and sustained expression of therapeutic proteins, such as apolipoprotein E (APOE), dramatically reducing plasma lipids in a model of hypercholesterolemia. Finally, we demonstrate that Apoa1-targeted fumarylacetoacetate hydrolase (FAH) can correct and rescue the severe metabolic liver disease hereditary tyrosinemia type I. In summary, we identify and validate Apoa1 as a novel integration site that supports durable transgene expression in the liver for gene therapy applications.
Graphical Abstract
Included in
Biochemical Phenomena, Metabolism, and Nutrition Commons, Biochemistry, Biophysics, and Structural Biology Commons, Biology Commons, Endocrinology, Diabetes, and Metabolism Commons, Genetic Phenomena Commons, Genetic Processes Commons, Medical Genetics Commons
