title: A piggyBac insertion disrupts Foxl2 expression that mimics BPES syndrome in mice.
abstract: Blepharophimosis, ptosis, epicanthus inversus syndrome (BPES) is an autosomal dominant genetic disorder characterized by small palpebral fissures and other craniofacial malformations, often with (type I) but could also without (type II) premature ovarian failure. While mutations of the forkhead transcription factor FOXL2 are associated with and likely be responsible for many BPES cases, how FOXL2 affects craniofacial development remain to be understood. Through a large-scale piggyBac (PB) insertion mutagenesis, we have identified a mouse mutant carrying a PB insertion ∼160 kb upstream of the transcription start site (TSS) of Foxl2. The insertion reduces, but not eliminates, the expression of Foxl2. This mutant, but not its revertant, displays BPES-like conditions such as midface hypoplasia, eyelid abnormalities and female subfertility. Further analysis indicates that the mutation does not affect mandible, but causes premature fusion of the premaxilla-maxilla suture, smaller premaxilla and malformed maxilla during midface development. We further identified an evolutionarily conserved fragment near the insertion site and observed enhancer activity of this element in tissue culture cells. Analyses using DNase I hypersensitivity assay and chromosome conformation capture assay in developing maxillary and periocular tissues suggest that the DNA region near the insertion site likely interacts with Foxl2 TSS. Therefore, this mutant presents an excellent animal model for mechanistic study of BPES and regulation of Foxl2.
authors.affiliation: State Key Laboratory of Genetic Engineering and National Center for International Research of Development and Disease, Institute of Developmental Biology and Molecular Medicine, Collaborative Innovation Center for Genetics and Development, School of Life Sciences, Fudan University, Shanghai 200433, China.
authors.forename: Fubiao
authors.initials: F
authors.lastname: Shi
authors.affiliation: State Key Laboratory of Genetic Engineering and National Center for International Research of Development and Disease, Institute of Developmental Biology and Molecular Medicine, Collaborative Innovation Center for Genetics and Development, School of Life Sciences, Fudan University, Shanghai 200433, China Howard Hughes Medical Institute, Department of Genetics, Yale University School of Medicine, New Haven, CT 06536, USA.
authors.forename: Sheng
authors.initials: S
authors.lastname: Ding
authors.affiliation: State Key Laboratory of Genetic Engineering and National Center for International Research of Development and Disease, Institute of Developmental Biology and Molecular Medicine, Collaborative Innovation Center for Genetics and Development, School of Life Sciences, Fudan University, Shanghai 200433, China.
authors.forename: Shimin
authors.initials: S
authors.lastname: Zhao
authors.affiliation: State Key Laboratory of Genetic Engineering and National Center for International Research of Development and Disease, Institute of Developmental Biology and Molecular Medicine, Collaborative Innovation Center for Genetics and Development, School of Life Sciences, Fudan University, Shanghai 200433, China Howard Hughes Medical Institute, Department of Molecular, Cellular, Developmental Biology, University of Colorado, Boulder, CO 80309, USA.
authors.forename: Min
authors.initials: M
authors.lastname: Han
authors.affiliation: State Key Laboratory of Genetic Engineering and National Center for International Research of Development and Disease, Institute of Developmental Biology and Molecular Medicine, Collaborative Innovation Center for Genetics and Development, School of Life Sciences, Fudan University, Shanghai 200433, China Department of Immunology, Duke University Medical Center, Durham, NC 27710, USA.
authors.forename: Yuan
authors.initials: Y
authors.lastname: Zhuang
authors.affiliation: State Key Laboratory of Genetic Engineering and National Center for International Research of Development and Disease, Institute of Developmental Biology and Molecular Medicine, Collaborative Innovation Center for Genetics and Development, School of Life Sciences, Fudan University, Shanghai 200433, China Howard Hughes Medical Institute, Department of Genetics, Yale University School of Medicine, New Haven, CT 06536, USA.
authors.forename: Tian
authors.initials: T
authors.lastname: Xu
authors.affiliation: State Key Laboratory of Genetic Engineering and National Center for International Research of Development and Disease, Institute of Developmental Biology and Molecular Medicine, Collaborative Innovation Center for Genetics and Development, School of Life Sciences, Fudan University, Shanghai 200433, China xiaohui_wu@fudan.edu.cn.
authors.forename: Xiaohui
authors.initials: X
authors.lastname: Wu
mesh_terms: D000818:Animals; D016569:Blepharophimosis / Q000235:genetics / Q000473:pathology*; D004251:DNA Transposable Elements; D004195:Disease Models, Animal; D000074842:Forkhead Box Protein L2; D051858:Forkhead Transcription Factors / Q000235:genetics* / Q000378:metabolism*; D006801:Humans; D008437:Maxilla / Q000254:growth & development / Q000473:pathology; D051379:Mice; D016254:Mutagenesis, Insertional; D012868:Skin Abnormalities / Q000235:genetics / Q000473:pathology*; D014564:Urogenital Abnormalities
chemical_list: D004251:DNA Transposable Elements; D000074842:Forkhead Box Protein L2; D051858:Forkhead Transcription Factors; C498893:Foxl2 protein, mouse