Identification of CTLA2A, DEFB29, WFDC15B, SERPINA1F and MUP19 as Novel Tissue-Specific Secretory Factors in Mouse

Abstract

Secretory factors in animals play an important role in communication between different cells, tissues and organs. Especially, the secretory factors with specific expression in one tissue may reflect important functions and unique status of that tissue in an organism. In this study, we identified potential tissue-specific secretory factors in the fat, muscle, heart, lung, kidney and liver in the mouse by analyzing microarray data from NCBI’s Gene Expression Omnibus (GEO) public repository and searching and predicting their subcellular location in GeneCards and WoLF PSORT, and then confirmed tissue-specific expression of the genes using semi-quantitative PCR reactions. With this approach, we confirmed 11 lung, 7 liver, 2 heart, 1 heart and muscle, 7 kidney and 2 adipose and liver-specific secretory factors. Among these genes, 1 lung-specific gene - CTLA2A (cytotoxic T lymphocyte-associated protein 2 alpha), 3 kidney-specific genes - SERPINA1F (serpin peptidase inhibitor, Clade A, member 1F), WFDC15B (WAP four-disulfide core domain 15B) and DEFB29 (defensin beta 29) and 1 liver-specific gene - MUP19 (major urinary protein 19) have not been reported as secretory factors. These genes were tagged with hemagglutinin at the 3’end and then transiently transfected to HEK293 cells. Through protein detection in cell lysate and media using Western blotting, we verified secretion of the 5 genes and predicted the potential pathways in which they may participate in the specific tissue through data analysis of GEO profiles. In addition, alternative splicing was detected in transcripts of CTLA2A and SERPINA1F and the corresponding proteins were found not to be secreted in cell culture media. Identification of novel secretory factors through the current study provides a new platform to explore novel secretory factors and a general direction for further study of these genes in the future.

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

“…Microarray database in the Gene Expression Omnibus (GEO), a public genomics data repository, was examined to determine intestine-specific genes among mucin ( MUC ) genes, as described in our previous reports [ 1 , 2 ]. In particular, GEO DataSet (GDS) 3142 for the mouse MUC genes and GDS3113 for the human MUC genes were obtained from the NCBI website, and gene expression profiles for eight tissues (small intestine, spleen, muscle, liver, brain, lung, kidney, and heart) were sorted out based on the normalized expression value.…”

“…Therefore, identification of tissue- and stage-specific genes and characterization of their promoters is important in order to promote transgene expression in a tissue- and stage-specific manner in genetically engineered animals for the elucidation of transgene function. To identify numerous tissue-specific genes in both mice and humans, comparative microarray database analysis based on the Gene Expression Omnibus (GEO) repository was previously used in our studies, and several novel tissue-specific genes were discovered [ 1 , 2 ]. In addition, our comparative analysis of the promoter region of tissue-specific genes has identified conserved cis -acting elements as a major aspect of tissue-specific regulation and expression [ 3 , 4 , 5 ].…”

Adipose‐Specific Expression, Developmental and Nutritional Regulation of the Gene‐Encoding Retinol‐Binding Protein 7 in Pigs

“…Tissue distribution of the RBP7 mRNA expression was investigated using a publicly available RNA‐seq dataset (accession number GSE77776) that was downloaded from the NCBI Gene Expression Omnibus (GEO; https://www.ncbi.nlm.nih.gov/geo/). The RNA‐seq data for seven different tissues (subcutaneous adipose tissue, heart, muscle, spleen, lung, liver, and kidney) from pigs ( n = 8) were sorted and analyzed as described in our previous reports (Song et al, ; Zhang et al, ).…”

“…ncbi.nlm.nih.gov/geo/). The RNA-seq data for seven different tissues (subcutaneous adipose tissue, heart, muscle, spleen, lung, liver, and kidney) from pigs (n = 8) were sorted and analyzed as described in our previous reports (Song et al, 2013;Zhang et al, 2015).…”

062 Identification of the MUC2 promoter as a strong promoter for intestinal gene expression through generation of transgenic quail expressing GFP in gut epithelial cells

“…Therefore, identification of tissue-and stage-specific genes and characterization of their promoters is important in order to promote transgene expression in a tissueand stage-specific manner in genetically engineered animals for the elucidation of transgene function. To identify numerous tissue-specific genes in both mice and humans, comparative microarray database analysis based on the Gene Expression Omnibus (GEO) repository was previously used in our studies, and several novel tissue-specific genes were discovered [1,2]. In addition, our comparative analysis of the promoter region of tissue-specific genes has identified conserved cis-acting elements as a major aspect of tissue-specific regulation and expression [3][4][5].…”