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Int J Mol Sci. 2017 Jan 19;18(1). pii: E196. doi: 10.3390/ijms18010196.

Identification of the MUC2 Promoter as a Strong Promoter for Intestinal Gene Expression through Generation of Transgenic Quail Expressing GFP in Gut Epithelial Cells.

Author information

1
Department of Animal Sciences, The Ohio State University, Columbus, OH 43210, USA. woodfint.1@osu.edu.
2
Department of Animal Sciences, The Ohio State University, Columbus, OH 43210, USA. chen.1930@osu.edu.
3
Department of Animal Sciences, The Ohio State University, Columbus, OH 43210, USA. ahn.134@osu.edu.
4
Department of Animal Sciences, The Ohio State University, Columbus, OH 43210, USA. suh.83@osu.edu.
5
Animal Biotechnology Division, National Institute of Animal Science, RDA, Wanju-gun, Jeonbuk 55365, Korea. hwangss@korea.kr.
6
Department of Animal Science and Technology, Sunchon National University, Suncheon 57922, Korea. rumen@sunchon.ac.kr.
7
Department of Animal Sciences, The Ohio State University, Columbus, OH 43210, USA. lee.2626@osu.edu.

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 of cis-acting elements in avian MUC2 gene 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 chicken MUC2 promoter 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 kb MUC2 promoter could retain the regulatory element to drive expression of target genes in intestinal tissues after hatching. This new transgene expression system, using the MUC2 promoter, will provide a new method of overexpressing target genes to study gene function in the avian intestine.

KEYWORDS:

Japanese quail; eGFP; intestine-specific; mucin 2; promoter; transgenic

PMID:
28106824
PMCID:
PMC5297827
DOI:
10.3390/ijms18010196
[Indexed for MEDLINE]
Free PMC Article

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