Crispr/cas Mediated Gene-targeting to Reduce Milk Allergens and Mastitis in Goats

Goat meat and milk is a major source of livelihood of small farmers. It is the only source of protein for many people in developing and underdeveloped countries. This industry is rapidly expanding in southern US due to its nutritional benefits, manageable operations, and influx of Africans, Hispanics and Asian population in recent years. Production losses occur for both dairy and meat farmers due to mammary gland infections (mastitis) affecting the health of both mother and the suckling kids, in addition to loss of marketable milk. Although, goat milk is superior to cow nutritionally and is used for infant formula in humans, presence of ?-lactoglobin (BLG), a known allergen for humans, makes it less desirable for human consumption. Therefore, there are two major scientific objectives of this project: the first is to use genome editing tools, specifically CRISPR/Cas system to generate a goat knockout for BLG gene (so as to eliminate human allergen in milk), that resembles a naturally occurring mutation with no foreign DNA. The secord is to establish and validate genome editing tools to create a transgenic goat expressing a peptidoglycan hydrolase enzyme (antibacterial protein) in mammary epithelial cells (expressed from the mammary-specific BLG gene, to prevent mastitis) for future investigation of the animals? ability to defend against a mastitis pathogen challenge.