title: Pancreatic cell tracing, lineage tagging and targeted genetic manipulations in multiple cell types using pancreatic ductal infusion of adeno-associated viral vectors and/or cell-tagging dyes.
abstract: Genetic manipulations, with or without lineage tracing for specific pancreatic cell types, are very powerful tools for studying diabetes, pancreatitis and pancreatic cancer. Nevertheless, the use of Cre/loxP systems to conditionally activate or inactivate the expression of genes in a cell type- and/or temporal-specific manner is not applicable to cell tracing and/or gene manipulations in more than one lineage at a time. Here we report a technique that allows efficient delivery of dyes for cell tagging into the mouse pancreas through the duct system, and that also delivers viruses carrying transgenes or siRNA under a specific promoter. When this technique is applied in genetically modified mice, it enables the investigator to perform either double lineage tracing or cell lineage tracing combined with gene manipulation in a second lineage. The technique requires <40 min.
authors.affiliation: Division of Pediatric Surgery, Children's Hospital of Pittsburgh, University of Pittsburgh School of Medicine, Pittsburgh, Pennsylvania, USA.
authors.forename: Xiangwei
authors.initials: X
authors.lastname: Xiao
authors.affiliation: Division of Pediatric Surgery, Children's Hospital of Pittsburgh, University of Pittsburgh School of Medicine, Pittsburgh, Pennsylvania, USA.
authors.forename: Ping
authors.initials: P
authors.lastname: Guo
authors.affiliation: Division of Pediatric Surgery, Children's Hospital of Pittsburgh, University of Pittsburgh School of Medicine, Pittsburgh, Pennsylvania, USA.
authors.forename: Krishna
authors.initials: K
authors.lastname: Prasadan
authors.affiliation: Division of Pediatric Surgery, Children's Hospital of Pittsburgh, University of Pittsburgh School of Medicine, Pittsburgh, Pennsylvania, USA.
authors.forename: Chiyo
authors.initials: C
authors.lastname: Shiota
authors.affiliation: Division of Pediatric Surgery, Children's Hospital of Pittsburgh, University of Pittsburgh School of Medicine, Pittsburgh, Pennsylvania, USA.
authors.forename: Lauren
authors.initials: L
authors.lastname: Peirish
authors.affiliation: Division of Pediatric Surgery, Children's Hospital of Pittsburgh, University of Pittsburgh School of Medicine, Pittsburgh, Pennsylvania, USA.
authors.forename: Shane
authors.initials: S
authors.lastname: Fischbach
authors.affiliation: Division of Pediatric Surgery, Children's Hospital of Pittsburgh, University of Pittsburgh School of Medicine, Pittsburgh, Pennsylvania, USA.
authors.forename: Zewen
authors.initials: Z
authors.lastname: Song
authors.affiliation: Division of Pediatric Surgery, Children's Hospital of Pittsburgh, University of Pittsburgh School of Medicine, Pittsburgh, Pennsylvania, USA.
authors.forename: Iljana
authors.initials: I
authors.lastname: Gaffar
authors.affiliation: Division of Pediatric Surgery, Children's Hospital of Pittsburgh, University of Pittsburgh School of Medicine, Pittsburgh, Pennsylvania, USA.
authors.forename: John
authors.initials: J
authors.lastname: Wiersch
authors.affiliation: Division of Pediatric Surgery, Children's Hospital of Pittsburgh, University of Pittsburgh School of Medicine, Pittsburgh, Pennsylvania, USA.
authors.forename: Yousef
authors.initials: Y
authors.lastname: El-Gohary
authors.affiliation: Division of Pediatrics, Children's Hospital of Pittsburgh, University of Pittsburgh School of Medicine, Pittsburgh, Pennsylvania, USA.
authors.forename: Sohail Z
authors.initials: SZ
authors.lastname: Husain
authors.affiliation: Division of Pediatric Surgery, Children's Hospital of Pittsburgh, University of Pittsburgh School of Medicine, Pittsburgh, Pennsylvania, USA.
authors.forename: George K
authors.initials: GK
authors.lastname: Gittes
mesh_terms: D000818:Animals; D019070:Cell Lineage; D004396:Coloring Agents / Q000008:administration & dosage*; D000229:Dependovirus / Q000235:genetics*; D004123:Dimethylamines / Q000008:administration & dosage; D005260:Female; D005821:Genetic Techniques; D005822:Genetic Vectors; D007813:Laparotomy; D008297:Male; D008810:Mice, Inbred C57BL; D008822:Mice, Transgenic; D010179:Pancreas / Q000166:cytology*; D010183:Pancreatic Ducts / Q000502:physiology / Q000601:surgery*; D011401:Promoter Regions, Genetic; D034741:RNA, Small Interfering; D014161:Transduction, Genetic; D019076:Transgenes
chemical_list: D004396:Coloring Agents; D004123:Dimethylamines; D034741:RNA, Small Interfering; C014518:dodecyldimethylamine oxide