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To produce melatonin\u2010enriched milk will benefit the consumers. In this study, a sheep bioreactor which generates melatonin\u2010enriched milk has been successfully developed by the technology that combined <jats:styled-content style=\"fixed-case\">CRISPR<\/jats:styled-content>\/Cas9 system and microinjection. The <jats:italic><jats:styled-content style=\"fixed-case\">AANAT<\/jats:styled-content><\/jats:italic> and <jats:italic><jats:styled-content style=\"fixed-case\">ASMT<\/jats:styled-content><\/jats:italic> were cloned from pineal gland of Dorper sheep (<jats:italic>Ovis aries<\/jats:italic>). The in vitro studies found that <jats:italic><jats:styled-content style=\"fixed-case\">AANAT<\/jats:styled-content><\/jats:italic> and <jats:italic><jats:styled-content style=\"fixed-case\">ASMT<\/jats:styled-content><\/jats:italic> were successfully transferred to the mammary epithelial cell lines and significantly increased melatonin production in the culture medium compared to the nontransgenic cell lines. In addition, the Cas9 <jats:styled-content style=\"fixed-case\">mRNA<\/jats:styled-content>, sg<jats:styled-content style=\"fixed-case\">RNA<\/jats:styled-content>, and the linearized vectors <jats:styled-content style=\"fixed-case\">pBC<\/jats:styled-content>1\u2010<jats:italic><jats:styled-content style=\"fixed-case\">AANAT<\/jats:styled-content><\/jats:italic> and <jats:styled-content style=\"fixed-case\">pBC<\/jats:styled-content>1\u2010<jats:italic><jats:styled-content style=\"fixed-case\">ASMT<\/jats:styled-content><\/jats:italic> were co\u2010injected into the cytoplasm of pronuclear embryos which were implanted into ewes by oviducts transferring. Thirty\u2010four transgenic sheep were generated with the transgenic positive rate being roughly 35% which were identified by Southern blot and sequencing. Seven carried transgenic <jats:italic><jats:styled-content style=\"fixed-case\">AANAT<\/jats:styled-content>,<\/jats:italic> two carried <jats:italic><jats:styled-content style=\"fixed-case\">ASMT<\/jats:styled-content>,<\/jats:italic> and 25 carried both of <jats:italic><jats:styled-content style=\"fixed-case\">AANAT<\/jats:styled-content><\/jats:italic> and <jats:italic><jats:styled-content style=\"fixed-case\">ASMT<\/jats:styled-content><\/jats:italic> genes. <jats:styled-content style=\"fixed-case\">RT<\/jats:styled-content>\u2010<jats:styled-content style=\"fixed-case\">PCR<\/jats:styled-content> and Western blot demonstrated that the lambs expressed these genes in their mammary epithelial cells and these animals produced melatonin\u2010enriched milk. This is the first report to show a functional <jats:italic><jats:styled-content style=\"fixed-case\">AANAT<\/jats:styled-content><\/jats:italic> and <jats:italic><jats:styled-content style=\"fixed-case\">ASMT<\/jats:styled-content><\/jats:italic> transgenic animal model which produce significantly high levels of melatonin milk compared to their wild\u2010type counterparts. The advanced technologies used in the study laid a foundation for generating large transgenic livestock, for example, the cows, which can produce high level of melatonin milk.<\/jats:p>","DOI":"10.1111\/jpi.12406","type":"journal-article","created":{"date-parts":[[2017,3,8]],"date-time":"2017-03-08T21:02:44Z","timestamp":1489006964000},"source":"Crossref","is-referenced-by-count":47,"title":["An <i><scp>AANAT<\/scp>\/<scp>ASMT<\/scp><\/i> transgenic animal model constructed with <scp>CRISPR<\/scp>\/Cas9 system serving as the mammary gland bioreactor to produce melatonin\u2010enriched milk in sheep"],"prefix":"10.1111","volume":"63","author":[{"given":"Teng","family":"Ma","sequence":"first","affiliation":[{"name":"National Engineering Laboratory for Animal Breeding Key Laboratory of Animal Genetics and Breeding of the Ministry of Agriculture Beijing Key Laboratory for Animal Genetic Improvement College of Animal Science and Technology China Agricultural University  Beijing 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