Enhancing the antibacterial activities of sow milk via site-specific knock-in of a lactoferrin gene in pigs using CRISPR
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LETTER TO THE EDITOR
Cell & Bioscience Open Access
Enhancing the antibacterial activities of sow milk via site‑specific knock‑in of a lactoferrin gene in pigs using CRISPR/Cas9 technology Xiaosong Han1, Yang Gao1, Guanglei Li1, Youcai Xiong1, Changzhi Zhao1, Jinxue Ruan1, Yunlong Ma1, Xinyun Li1,2, Changchun Li1,2, Shuhong Zhao1,2 and Shengsong Xie1,2*
Abstract Colostrum quality is a vital factor in mortality and growth performance for piglets. Lactoferrin is an immuno-active milk protein that contributes to the formation of a protective layer above intestinal mucosa, possesses the antibacterial and antiviral activities that are favorable for piglet development. However, there is a notable reduction in lactoferrin in sow milk during lactation after the first few days, which causes many piglets to fail to ingest enough colostrum thereby leading to an increase in piglet mortality. In this study, we successfully constructed genome-edited LargeWhite pigs with marker-free site-specific knock-in of lactoferrin gene in the 3′-end of Casein alpha-s1 via CRISPR/ Cas9 mediated homologous recombination. Thus, the lactoferrin protein can be expressed in the mammary gland in the control of Casein alpha-s1 promoter. As expected, the lactoferrin protein in genetically modified pigs sustained high expression in both colostrum and milk when compared with wild-type pigs. Moreover, the bacterial plate assay indicated that the milk from genetically modified pigs showed bacteriostatic effects when compared with control pigs. Taken together, our study demonstrated that the milk from genetically modified pigs had antibacterial activity which may reduce the costs of veterinary drug and improve the surviving rate of piglets, which is promising for pig breeding. Keywords: Pig, Lactoferrin, Colostrum, CRISPR/Cas9, Homologous recombination Dear Editor, Due to the lack of a fully developed immune system, newborn piglets are susceptible to pathogenic bacteria, thus causing billions of dollars in annual global losses. Sow milk, especially colostrum, containing abundant immune active compounds, plays a key role in piglet thermoregulation, acquisition of passive immunity, and intestinal development [1]. Colostrum quality is a vital *Correspondence: [email protected] 1 Key Laboratory of Agricultural Animal Genetics, Breeding and Reproduction of Ministry of Education & Key Lab of Swine Genetics and Breeding of Ministry of Agriculture and Rural Affairs, Huazhong Agricultural University, Wuhan 430070, People’s Republic of China Full list of author information is available at the end of the article
factor in mortality and growth performance for piglets. Lactoferrin (LF) is an immune-active milk protein that contributes to the formation of a protective layer above intestinal mucosa, possesses antibacterial and antiviral properties [2], therefore, piglets suffer less from intestinal inflammation or diarrhea. However, the secretion volume for porcine lactoferrin (pLF) decreases in sow milk along with the lactation. Previous studies have shown th
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