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

Overview

Journal Nat Protoc

Specialties Biology
Pathology
Science

Date 2014 Oct 31

PMID 25356582

Citations 46

Authors

Xiangwei Xiao

Ping Guo

Krishna Prasadan

Chiyo Shiota

Lauren Peirish

Shane Fischbach

Zewen Song

Iljana Gaffar

John Wiersch

Yousef El-Gohary

Sohail Z Husain

George K Gittes

Affiliations

Soon will be listed here.

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.

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