ΦC31 Integrase Mediates Efficient Site-Specific Integration in Sheep Fibroblasts

• Published in: Bioscience, biotechnology, and biochemistry Vol. 76; no. 11; pp. 2093 - 2095
• Main Authors: NI, Wei, HU, Shengwei, QIAO, Jun, WANG, Yuanzhi, SHI, Huijun, WANG, Yong, HE, Zhirui, LI, Guozhong, CHEN, Chuangfu
• Format: Journal Article
• Language: English
• Published: Tokyo Japan Society for Bioscience, Biotechnology, and Agrochemistry 23.11.2012; Japan Society for Bioscience Biotechnology and Agrochemistry
• Subjects: Animals; Attachment Sites, Microbiological; Biological and medical sciences; Fibroblasts - metabolism; Fundamental and applied biological sciences. Psychology; Gene Transfer Techniques; Genomics; Green Fluorescent Proteins - genetics; Integrases - metabolism; pseudo attP sites; Sheep; sheep fibroblasts; specific integration; Transgenes - genetics; ΦC31 integrase; Site specificity; Vertebrata; Mammalia; specific integration; Sheep; Artiodactyla; Fibroblast; sheep fibroblasts; ΦC31 integrase; Ungulata; pseudo attP sites
• ISSN: 0916-8451; 1347-6947
• DOI: 10.1271/bbb.120439

ΦC31 integrase, a site-specific recombinase, can effectively mediate the integration of foreign genes bearing an attB sequence into pseudo attP sites of genomes in human, mouse, and Drosophila cells. In this study, we measured ΦC31 integrase-mediated homologous recombination between attB and pseudo attP sites in sheep cells. The integration efficiency of the EGFP expression cassette with the attB sequence increased at least 2-fold in sheep fibroblasts. Three pseudo-attP sites were identified in the sheep genome, located in the intergenic regions on chromosomes 4, 13, and 7 respectively. Moreover, the transgene that was integrated at the three pseudo attP sites exhibited high levels of expression. Our study indicates that the ΦC31 integrase system provides an efficient integration tool for genetic engineering of the sheep genome.