{"database":"GEO","file_versions":[{"headers":{"Content-Type":["application/json"]},"body":{"files":{"Other":["ftp://ftp.ncbi.nlm.nih.gov/geo/series/GSE308nnn/GSE308034/"]},"type":"primary"},"statusCode":"OK","statusCodeValue":200}],"scores":null,"additional":{"omics_type":["Other"],"species":["Mus musculus"],"gds_type":["Other"],"full_dataset_link":["https://www.ncbi.nlm.nih.gov/geo/query/acc.cgi?acc=GSE308034"],"repository":["GEO"],"entry_type":["GSE"],"additional_accession":[]},"is_claimable":false,"name":"Highly efficient and low-mosaicism piggyBac transgenesis platform for rapid founder phenotyping [Amplicon-seq]","description":"Pronuclear microinjection is the most widely used method for producing transgenic (Tg) animals. Because its efficiency is typically <20%, phenotypic characterization of Tg animals is usually performed from the next generation (F1) onwards. Conventional Cre-loxP-based conditional knockout (cKO) mouse production requires generating two genetically modified strains and multiple rounds of breeding before cKO mice are available for analysis. Here, we optimized a piggyBac transposon-based method of Tg mouse production and established conditions under which nearly all founder (F0) embryos are transgenic. Using a single-cell RNA sequencing-based strategy, we characterized mosaicism in F0 embryos and demonstrated that piggyBac-mediated transgene integration occurs early in embryonic development. We also achieved ~70% efficiency in generating bacterial-artificial-chromosome-Tg mice. By combining this method with genome editing, we developed a strategy for tissue-specific-knockout phenotyping in the F0 generation. Our findings establish a highly efficient piggyBac-based platform that expands experimental options for Tg animal production by supporting rapid F0-based phenotypic assessment while also facilitating efficient generation of F0 animals for subsequent breeding.","dates":{"publication":"2026/06/11"},"accession":"GSE308034","cross_references":{"GSM":["GSM9236193","GSM9236194","GSM9236191","GSM9236192","GSM9236190","GSM9236201","GSM9236202","GSM9236199","GSM9236189","GSM9236200","GSM9236197","GSM9236198","GSM9236195","GSM9236196","GSM9236207","GSM9236205","GSM9236206","GSM9236203","GSM9236204"],"GPL":["32071"],"GSE":["308034"],"taxon":["Mus musculus"]}}