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.
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.
复制标题
DOI：
10.1038/nprot.2014.183
复制
发表时间：
2014-12
期刊：
影响因子：
14.8
通讯作者：
Gittes GK
中科院分区：
文献类型：
--
作者：
Xiao X;Guo P;Prasadan K;Shiota C;Peirish L;Fischbach S;Song Z;Gaffar I;Wiersch J;El-Gohary Y;Husain SZ;Gittes GK
基因操作，有或没有特定胰腺细胞类型的谱系追踪，是研究糖尿病，胰腺炎和胰腺癌的非常强大的工具。然而，使用Cre/loxP系统以细胞类型特异性和/或时间特异性方式有条件地激活或抑制基因表达不适用于一次在多于一个谱系中的细胞追踪和/或基因操作。在这里，我们报告了一种技术，允许有效地交付染料的细胞标记到小鼠胰腺通过导管系统，也提供了一个特定的启动子下携带转基因或siRNA的病毒。当这种技术应用于转基因小鼠时，它使研究人员能够进行双谱系追踪或细胞谱系追踪结合第二谱系中的基因操作。该技术需要<40 min。
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.