{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2025,10,10]],"date-time":"2025-10-10T18:59:52Z","timestamp":1760122792151,"version":"3.37.3"},"reference-count":54,"publisher":"Wiley","issue":"8","license":[{"start":{"date-parts":[[2018,10,2]],"date-time":"2018-10-02T00:00:00Z","timestamp":1538438400000},"content-version":"vor","delay-in-days":1,"URL":"http:\/\/onlinelibrary.wiley.com\/termsAndConditions#vor"}],"funder":[{"DOI":"10.13039\/501100001809","name":"National Natural Science Foundation of China","doi-asserted-by":"publisher","award":["31772836","31571504"],"award-info":[{"award-number":["31772836","31571504"]}],"id":[{"id":"10.13039\/501100001809","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":["onlinelibrary.wiley.com"],"crossmark-restriction":true},"short-container-title":["Environ and Mol Mutagen"],"published-print":{"date-parts":[[2018,10]]},"abstract":"<jats:p>Saturation mutagenesis of all endogenous genes within the mouse genome remains a challenging task, although a plenty of gene\u2010editing approaches are available for this purpose. Here, a poly(A)\u2010trap vector was generated for insertion mutagenesis in mouse embryonic stem (mES) cells. This vector contains an expression cassette of neomycin (Neo)\u2010resistant gene lacking a poly(A) signal and flanked by two inverted terminal repeats of the <jats:italic>Sleeping Beauty<\/jats:italic> (SB) transposon. The whole poly(A)\u2010trap cassette can transpose into target TA dinucleotides, properly splice with endogenous genes and effectively interrupt the transcription of trapped genes in mES cells after transient induction of SB expression by doxycycline (DOX)\u2010treatment at 1 \u03bcg\/ml, leading to the formation of multiple geneticin (G418)\u2010resistant cell clones. In the first round of mutation screening, we identified six transposition events from 23 cell clones, including four inserted into an endogenous gene and two landed between endogenous genes. The abilities of self\u2010renewal, totipotency, genetic stability and differentiation of <jats:italic>syngap1<\/jats:italic><jats:sup><jats:italic>+\/\u2212<\/jats:italic><\/jats:sup> cells were not affected by DOX\u2010treatment and G418\u2010selection. These findings suggest that this SB transposon\u2010mediated poly(A)\u2010trap vector can be used as an alternative tool for a large\u2010scale screening of mES cells with a gene mutation and for further generation of mutant mouse strains. Environ. Mol. Mutagen. 59:687\u2013697, 2018. \u00a9 2018 Wiley Periodicals, Inc.<\/jats:p>","DOI":"10.1002\/em.22234","type":"journal-article","created":{"date-parts":[[2018,10,3]],"date-time":"2018-10-03T06:47:22Z","timestamp":1538549242000},"page":"687-697","update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":1,"title":["<i>Sleeping beauty<\/i> transposon\u2010mediated poly(A)\u2010trapping and insertion mutagenesis in mouse embryonic stem cells"],"prefix":"10.1002","volume":"59","author":[{"ORCID":"https:\/\/orcid.org\/0000-0003-1746-9000","authenticated-orcid":false,"given":"Yi","family":"Zhao","sequence":"first","affiliation":[{"name":"Department of Genetics, School of Basic Medical Sciences Wuhan University  Wuhan Hubei China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Guili","family":"Song","sequence":"additional","affiliation":[{"name":"State Key Laboratory of Freshwater Ecology and Biotechnology Institute of 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