Targeting Human ?-Lactalbumin Gene Insertion into the Goat ?-Lactoglobulin Locus by TALEN-Mediated Homologous Recombination.

ABSTRACT: Special value of goat milk in human nutrition and well being is associated with medical problems of food allergies which are caused by milk proteins such as ?-lactoglobulin (BLG). Here, we employed transcription activator-like effector nuclease (TALEN)-assisted homologous recombination in goat fibroblasts to introduce human ?-lactalbumin (hLA) genes into goat BLG locus. TALEN-mediated targeting enabled isolation of colonies with mono- and bi-allelic transgene integration in up to 10.1% and 1.1%, respectively, after selection. Specifically, BLG mRNA levels were gradually decreasing in both mo- and bi-allelic goat mammary epithelial cells (GMECs) while hLA demonstrated expression in GMECs in vitro. Gene-targeted fibroblast cells were efficiently used in somatic cell nuclear transfer, resulting in production of hLA knock-in goats directing down-regulated BLG expression and abundant hLA secretion in animal milk. Our findings provide valuable background for animal milk optimization and expedited development for agriculture and biomedicine.

SUBMITTER: Zhu H 

PROVIDER: S-EPMC4892491 | biostudies-literature | 2016

REPOSITORIES: biostudies-literature