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/SRR263/022/SRR26368222/SRR26368222_1.fastq.gzftp://ftp.sra.ebi.ac.uk/vol1/fastq/SRR263/076/SRR26368476/SRR26368476_1.fastq.gzftp://ftp.sra.ebi.ac.uk/vol1/fastq/SRR263/099/SRR26368099/SRR26368099_2.fastq.gzftp://ftp.sra.ebi.ac.uk/vol1/fastq/SRR263/040/SRR26368240/SRR26368240_1.fastq.gzftp://ftp.sra.ebi.ac.uk/vol1/fastq/SRR263/094/SRR26368494/SRR26368494_1.fastq.gzftp://ftp.sra.ebi.ac.uk/vol1/fastq/SRR263/019/SRR26368219/SRR26368219_1.fastq.gzftp://ftp.sra.ebi.ac.uk/vol1/fastq/SRR263/061/SRR26368461/SRR26368461_1.fastq.gzftp://ftp.sra.ebi.ac.uk/vol1/fastq/SRR263/092/SRR26351392/SRR26351392.fastq.gzprimaryOK200GenomicsUniversity of Pennsylvaniahttps://www.ebi.ac.uk/ena/browser/view/PRJNA1026949xref:EuropePMC:PMC10800763The c.1222C>T (p.Arg408Trp) phenylalanine hydroxylase (PAH) variant is the most frequent cause of phenylketonuria (PKU), an autosomal recessive disorder characterized by accumulation of blood phenylalanine (Phe) to neurotoxic levels. Here we devised a therapeutic base editing strategy to correct the variant, using prime-edited hepatocyte cell lines engineered with the c.1222C>T variant to screen a variety of adenine base editors and guide RNAs in vitro, followed by assessment in c.1222C>T humanized mice in vivo. We found that upon delivery of a selected adenine base editor mRNA/guide RNA combination into mice via lipid nanoparticles (LNPs), there was sufficient PAH editing in the liver to fully normalize blood Phe levels within 48 hours. This work establishes the viability of a base editing strategy to correct the most common pathogenic variant found in individuals with the most common inborn error of metabolism, albeit with potential limitations compared to other genome editing approaches.ENAfalseA base editing strategy using mRNA-LNPs for in vivo correction of the most frequent phenylketonuria variant2023-10-282023-10-28PRJNA1026949