Wistar Institute View Institution's Website 4 articles published in JoVE Immunology and Infection Gene Editing of Primary Rhesus Macaque B Cells Harald Hartweger1, Rajeev Gautam2, Yoshiaki Nishimura2, Fabian Schmidt3,5, Kai-Hui Yao1, Amelia Escolano1,6, Mila Jankovic1, Malcolm A. Martin2, Michel C. Nussenzweig1,4 1Laboratory of Molecular Immunology, The Rockefeller University, 2Laboratory of Molecular Microbiology, National Institute of Allergy and Infectious Diseases, National Institutes of Health, 3Laboratory of Retrovirology, The Rockefeller University, 4Howard Hughes Medical Institute, The Rockefeller University, 5Laboratory of Applied Virology and Precision Medicine, King Abdullah University of Science and Technology (KAUST), 6Vaccine and Immunotherapy Center, Wistar Institute We present a method for culturing and gene editing primary rhesus macaque B cells using CRISPR/Cas9 and recombinant adeno-associated virus serotype 6 for the study of B cell therapies. Cancer Research A Melanoma Patient-Derived Xenograft Model Min Xiao1, Vito W. Rebecca1, Meenhard Herlyn1 1Wistar Institute Patient-derived xenograft (PDX) models more robustly recapitulate melanoma molecular and biological features and are more predictive of therapy response compared to traditional plastic tissue culture-based assays. Here we describe our standard operating protocol for the establishment of new PDX models and the characterization/experimentation of existing PDX models. Medicine Initiation of Metastatic Breast Carcinoma by Targeting of the Ductal Epithelium with Adenovirus-Cre: A Novel Transgenic Mouse Model of Breast Cancer Melanie R. Rutkowski*1, Michael J. Allegrezza*1, Nikolaos Svoronos1, Amelia J. Tesone1, Tom L. Stephen1, Alfredo Perales-Puchalt1, Jenny Nguyen1, Paul J. Zhang2, Steven N. Fiering3, Julia Tchou4,5,6, Jose R. Conejo-Garcia1 1Tumor Microenvironment and Metastasis Program, Wistar Institute, 2Department of Pathology and Lab Medicine, Perelman School of Medicine, University of Pennsylvania, 3Department of Microbiology and Immunology and Department of Genetics, Geisel School of Medicine at Dartmouth, 4Division of Endocrine and Oncologic Surgery, Department of Surgery, Perelman School of Medicine, University of Pennsylvania, 5Rena Rowan Breast Center, Abramson Cancer Center, University of Pennsylvania, 6Center for Advanced Medicine, University of Pennsylvania Activation of latent mutations with adenovirus-Cre into the mammary ductal system results in a clinically relevant metastatic breast cancer. Incorporation of a YFP promoter allows tracking of distal metastatic tumor cells. This model is useful to study latent metastasis, anti-tumor immunity, and for designing novel immunotherapies to treat breast cancer. Bioengineering The Three-Dimensional Human Skin Reconstruct Model: a Tool to Study Normal Skin and Melanoma Progression Ling Li1, Mizuho Fukunaga-Kalabis1, Meenhard Herlyn1 1Molecular and Cellular Oncogenesis Program, The Wistar Institute In this report, we describe the three-dimensional skin reconstruct model which mimics human skin in architecture and composition. Melanocyte physiology, melanoma progression and the fate of dermal stem cells have been investigated using the skin reconstruct model. The model is also useful as a preclinical tool for drug assessment.