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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Acknowledgements
This work was supported, in whole or in part, by the Cochrane-Weber endowed Fund in Diabetes Research (X.X., NO19831) and the US National Institutes of Health (G.K.G., R01 DK098196).
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The concept of the study was designed by X.X. Surgery was developed and performed by X.X., P.G. and G.K.G. Viruses were produced by P.G. Surgery photographs were taken by K.P. Data were collected by X.X., P.G., K.P., C.S., L.P., S.F., I.G., Y.E.G., S.Z.H. and J.W. X.X., S.Z.H. and G.K.G. wrote the article.
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Xiao, X., Guo, P., Prasadan, K. et al. 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. Nat Protoc 9, 2719–2724 (2014). https://doi.org/10.1038/nprot.2014.183
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DOI: https://doi.org/10.1038/nprot.2014.183
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