title: Identification of the MUC2 Promoter as a Strong Promoter for Intestinal Gene Expression through Generation of Transgenic Quail Expressing GFP in Gut Epithelial Cells.
abstract: Identification of tissue- and stage-specific gene promoters is valuable for delineating the functional roles of specific genes in genetically engineered animals. Here, through the comparison of gene expression in different tissues by analysis of a microarray database, the intestinal specificity of mucin 2 (MUC2) expression was identified in mice and humans, and further confirmed in chickens by RT-PCR (reverse transcription-PCR) analysis. An analysis ofcis-acting elements in avianMUC2gene promoters revealed conservation of binding sites, within a 2.9 kb proximal promoter region, for transcription factors such as caudal type homeobox 2 (CDX2), GATA binding protein 4 (GATA4), hepatocyte nuclear factor 4 α (HNF4A), and transcription factor 4 (TCF4) that are important for maintaining intestinal homeostasis and functional integrity. By generating transgenic quail, we demonstrated that the 2.9 kb chickenMUC2promoter could drive green fluorescent protein (GFP) reporter expression exclusively in the small intestine, large intestine, and ceca. Fluorescence image analysis further revealed GFP expression in intestine epithelial cells. The GFP expression was barely detectable in the embryonic intestine, but increased during post-hatch development. The spatiotemporal expression pattern of the reporter gene confirmed that the 2.9 kbMUC2promoter could retain the regulatory element to drive expression of target genes in intestinal tissues after hatching. This new transgene expression system, using theMUC2promoter, will provide a new method of overexpressing target genes to study gene function in the avian intestine.
authors.affiliation: Department of Animal Sciences, The Ohio State University, Columbus, OH 43210, USA. woodfint.1@osu.edu.
authors.forename: Rachel M
authors.initials: RM
authors.lastname: Woodfint
authors.affiliation: Department of Animal Sciences, The Ohio State University, Columbus, OH 43210, USA. chen.1930@osu.edu.
authors.forename: Paula R
authors.initials: PR
authors.lastname: Chen
authors.affiliation: Department of Animal Sciences, The Ohio State University, Columbus, OH 43210, USA. ahn.134@osu.edu.
authors.forename: Jinsoo
authors.initials: J
authors.lastname: Ahn
authors.affiliation: Department of Animal Sciences, The Ohio State University, Columbus, OH 43210, USA. suh.83@osu.edu.
authors.forename: Yeunsu
authors.initials: Y
authors.lastname: Suh
authors.affiliation: Animal Biotechnology Division, National Institute of Animal Science, RDA, Wanju-gun, Jeonbuk 55365, Korea. hwangss@korea.kr.
authors.forename: Seongsoo
authors.initials: S
authors.lastname: Hwang
authors.affiliation: Department of Animal Science and Technology, Sunchon National University, Suncheon 57922, Korea. rumen@sunchon.ac.kr.
authors.forename: Sang Suk
authors.initials: SS
authors.lastname: Lee
authors.affiliation: Department of Animal Sciences, The Ohio State University, Columbus, OH 43210, USA. lee.2626@osu.edu.
authors.forename: Kichoon
authors.initials: K
authors.lastname: Lee
mesh_terms: D000818:Animals; D030801:Animals, Genetically Modified; D001665:Binding Sites; D015153:Blotting, Western; D002645:Chickens; D004847:Epithelial Cells / Q000378:metabolism*; D005786:Gene Expression Regulation; D049452:Green Fluorescent Proteins / Q000378:metabolism*; D006801:Humans; D007422:Intestines / Q000166:cytology*; D051379:Mice; D055262:Mucin-2 / Q000235:genetics* / Q000378:metabolism; D020411:Oligonucleotide Array Sequence Analysis; D009928:Organ Specificity / Q000235:genetics; D011401:Promoter Regions, Genetic; D011784:Quail; D015203:Reproducibility of Results; D014018:Tissue Distribution
chemical_list: C524625:MUC2 protein, human; C524626:Muc2 protein, mouse; D055262:Mucin-2; D049452:Green Fluorescent Proteins
keywords: Japanese quail; eGFP; intestine-specific; mucin 2; promoter; transgenic