摘要：SummaryAmyotrophic lateral sclerosis/frontotemporal dementia (ALS/FTD) is a fatal neurodegenerative disorder, and continued innovation is needed for improved understanding and for developing therapeutics. We have created next-generationgenomically humanizedknockin mouse models, by replacing the mouse genomic region ofSod1,Tardbp(TDP-43), andFus, with their human orthologs, preserving human protein biochemistry and splicing with exons and introns intact. We establish a new standard of large knockin allele quality control, demonstrating the utility of indirect capture for enrichment of a genomic region of interest followed by Oxford Nanopore sequencing. Extensive analysis shows that homozygous humanized animals only express human protein at endogenous levels. Characterization of humanized FUS animals showed that they are phenotypically normal throughout their lifespan. These humanized strains are vital for preclinical assessment of interventions and serve as templates for the addition of coding or non-coding human ALS/FTD mutations to dissect disease pathomechanisms, in a physiological context.Graphical abstractDisplay OmittedHighlights•Replacement in mice ofSod1,Tardbp, andFuswith orthologous human genomic sequence•Thorough allele QC of knockin alleles using targeted long-read sequencing•Mice are viable, fertile, and provide refined tools for ALS/FTD research•Aging ofhFUSmice reveals no overt phenotype and minimal transcriptomic disruptionNeurogenetics; Neuroscience; Model organism
