title: Screening pancreatic oncogenes in zebrafish using the Gal4/UAS system.
abstract: Pancreatic cancer is a genetic disease in which somatic mutations in the KRAS proto-oncogene are detected in a majority of tumors. KRAS mutations represent an early event during pancreatic tumorigenesis, crucial for cancer initiation and progression. Recent studies, including comprehensive sequencing of the pancreatic cancer exome, have implicated the involvement of a number of additional core signaling pathways during pancreatic tumorigenesis. Improving our understanding of genetic interactions between KRAS and these additional pathways represents a critical challenge, as these interactions may provide novel opportunities for diagnosis and treatment. However, studying these interactions requires the expression of multiple transgenes in relevant cell types, an effort that has proven very difficult to achieve using gene targeted mice and is also technically challenging in zebrafish. Based on the ability of the Gal4 transcriptional activator to drive the expression of multiple transgenes under regulation of UAS (upstream activator sequence) regulatory elements, the Gal4/UAS system represents an attractive strategy for the study of genetic interactions. In this chapter, we review our experience using the Gal4/UAS system to model KRAS-initiated pancreatic cancer in zebrafish, as well as our early efforts using this system to study the influence of other cooperating oncogenes. We also describe techniques used to identify and characterize pancreatic tumors in adult transgenic fish.
authors.affiliation: Department of Surgery and McKusick-Nathans Institute of Genetic Medicine, Johns Hopkins University School of Medicine, Baltimore, Maryland, USA.
authors.forename: Shu
authors.initials: S
authors.lastname: Liu
authors.forename: Steven D
authors.initials: SD
authors.lastname: Leach
mesh_terms: D000818:Animals; D030801:Animals, Genetically Modified; D004268:DNA-Binding Proteins / Q000235:genetics / Q000378:metabolism; D004625:Embryo, Nonmammalian; D004742:Enhancer Elements, Genetic; D015972:Gene Expression Regulation, Neoplastic; D017930:Genes, Reporter; D049452:Green Fluorescent Proteins / Q000032:analysis / Q000235:genetics / Q000378:metabolism; D006801:Humans; D007150:Immunohistochemistry; D008495:Medical Oncology / Q000379:methods*; D008845:Microinjections / Q000379:methods*; D008867:Microtomy; D010179:Pancreas / Q000378:metabolism / Q000473:pathology*; D010190:Pancreatic Neoplasms / Q000235:genetics* / Q000378:metabolism / Q000473:pathology; D010957:Plasmids; D000090063:Proto-Oncogene Mas; D011518:Proto-Oncogene Proteins / Q000235:genetics / Q000378:metabolism*; D016283:Proto-Oncogene Proteins p21(ras); D029701:Saccharomyces cerevisiae Proteins / Q000235:genetics / Q000378:metabolism; D016610:Tissue Embedding / Q000379:methods*; D014157:Transcription Factors / Q000235:genetics / Q000378:metabolism; D019076:Transgenes; D015027:Zebrafish / Q000196:embryology / Q000235:genetics* / Q000378:metabolism; D018631:ras Proteins / Q000235:genetics / Q000378:metabolism*
chemical_list: D004268:DNA-Binding Proteins; C052130:GAL4 protein, S cerevisiae; C117307:KRAS protein, human; C000715470:MAS1 protein, human; D000090063:Proto-Oncogene Mas; D011518:Proto-Oncogene Proteins; D029701:Saccharomyces cerevisiae Proteins; D014157:Transcription Factors; C485184:enhanced green fluorescent protein; D049452:Green Fluorescent Proteins; D016283:Proto-Oncogene Proteins p21(ras); D018631:ras Proteins