Generating a transgenic mouse line stably expressing human MHC surface antigen from a HAC carrying multiple genomic BACs

• Published in: Chromosoma Vol. 124; no. 1; pp. 107 - 118
• Main Authors: Hasegawa, Yoshinori, Ishikura, Tomoyuki, Hasegawa, Takanori, Watanabe, Takashi, Suzuki, Junpei, Nakayama, Manabu, Okamura, Yoshiaki, Okazaki, Tuneko, Koseki, Haruhiko, Ohara, Osamu, Ikeno, Masashi, Masumoto, Hiroshi
• Format: Journal Article
• Language: English
• Published: Berlin/Heidelberg Springer Berlin Heidelberg 01.03.2015; Springer Nature B.V
• Subjects: Animal Genetics and Genomics; Animals; Biochemistry; Biomedical and Life Sciences; Cell Biology; CHO Cells; Chromosomes, Artificial, Bacterial; Chromosomes, Artificial, Human; Cricetulus; Developmental Biology; Eukaryotic Microbiology; Gene Expression Regulation; Genome; HLA-DR Antigens - genetics; Human Genetics; Humans; Life Sciences; Mice; Mice, Transgenic - genetics; Organ Specificity; Research Article; Transgenes; Major Histocompatibility Complex; Chinese Hamster Ovary Cell; Mouse Embryonic Stem Cell; Embryonic Stem Cell; Embryonic Stem Cell Line
• ISSN: 0009-5915; 1432-0886
• DOI: 10.1007/s00412-014-0488-3

The human artificial chromosome (HAC) vector is a promising tool to improve the problematic suppression and position effects of transgene expression frequently seen in transgenic cells and animals produced by conventional plasmid or viral vectors. We generated transgenic mice maintaining a single HAC vector carrying two genomic bacterial artificial chromosomes (BACs) from human HLA-DR loci (DRA and DRB1). Both transgenes on the HAC in transgenic mice exhibited tissue-specific expression in kidney, liver, lung, spleen, lymph node, bone marrow, and thymus cells in RT-PCR analysis. Stable functional expression of a cell surface HLA-DR marker from both transgenes, DRA and DRB1 on the HAC, was detected by flow cytometric analysis of splenocytes and maintained through at least eight filial generations. These results indicate that the de novo HAC system can allow us to manipulate multiple BAC transgenes with coordinated expression as a surface antigen through the generation of transgenic animals.