Rapid functional dissection of genetic networks via tissue-specific transduction and RNAi in mouse embryos.
Using ultrasound-guided in utero infections of fluorescently traceable lentiviruses carrying RNAi or Cre recombinase into mouse embryos, we have demonstrated noninvasive, highly efficient selective transduction of surface epithelium, in which progenitors stably incorporate and propagate the desired genetic alterations. We achieved epidermal-specific infection using small generic promoters of existing lentiviral short hairpin RNA libraries, thus enabling rapid assessment of gene function as well as complex genetic interactions in skin morphogenesis and disease in vivo. We adapted this technology to devise a new quantitative method for ascertaining whether a gene confers a growth advantage or disadvantage in skin tumorigenesis. Using alpha1-catenin as a model, we uncover new insights into its role as a widely expressed tumor suppressor and reveal physiological interactions between Ctnna1 and the Hras1-Mapk3 and Trp53 gene pathways in regulating skin cell proliferation and apoptosis. Our study illustrates the strategy and its broad applicability for investigations of tissue morphogenesis, lineage specification and cancers.
Beronja
Slobodan
S
Howard Hughes Medical Institute, Laboratory of Mammalian Cell Biology and Development, The Rockefeller University, New York, New York, USA.
Livshits
Geulah
G
Williams
Scott
S
Fuchs
Elaine
E
D000818:Animals; D051177:Catenins / Q000235:genetics; D049109:Cell Proliferation; D004622:Embryo, Mammalian; D004817:Epidermis; D005240:Feasibility Studies; D005822:Genetic Vectors; D019426:Integrases / Q000235:genetics; D016086:Lentivirus / Q000235:genetics; D051379:Mice; D009154:Mutation; D009928:Organ Specificity; D034622:RNA Interference; D014161:Transduction, Genetic; D014465:Ultrasonics
D051177:Catenins; C045073:Cre recombinase; D019426:Integrases