Nebraska Center for Virology 1 article published in JoVE Biochemistry A Double Humanized BLT-mice Model Featuring a Stable Human-Like Gut Microbiome and Human Immune System Lance Daharsh1,2, Jianshui Zhang1,2, Amanda Ramer-Tait3, Qingsheng Li1,2 1Nebraska Center for Virology, 2School of Biological Sciences, University of Nebraska-Lincoln, 3Department of Food Science and Technology, University of Nebraska-Lincoln We describe a novel method for generating double humanized BLT-mice that feature a functional human immune system and a stable engrafted human-like gut microbiome. This protocol can be followed without the need for germ-free mice or gnotobiotic facilities.