Generation and validation of a floxed FosB mouse line
Generation and validation of a floxed FosB mouse line
复制标题
floxed FosB 小鼠品系的生成和验证
DOI：
10.1101/179309
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发表时间：
2017
期刊：
影响因子：
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通讯作者：
Nestler Eric J.
中科院分区：
文献类型：
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作者：
Ohnishi Yoshinori N.;Eagle Andrew L.;Ohnishi Yoko H.;Cahill Michael E.;Wirtz Alexis J.;Robison Alfred J.;Nestler Eric J.
FosB基因的表达已经在许多领域使用各种工具进行了广泛的研究。然而，先前的技术具有各种警告，来自潜在的脱靶效应（例如，FosB、ΔFosB或JunD的显性负突变体（称为ΔJunD）的过表达）或混淆发育影响（例如，组成型FosB敲除小鼠）。因此，我们试图建立一个floxed FosB小鼠系，它将允许FosB基因的真正沉默，同时具有空间和时间控制。在这里，我们详细介绍了克隆策略，生产和验证的thefloxed FosB小鼠。我们证明了育种和基因分型的方法，并表明病毒介导的Cre重组酶在一个有针对性的，离散的大脑区域中的表达消融thefosB基因inhibited，但不是野生型小鼠的表达。因此，这里展示的floxed FosB小鼠代表了继续研究这一关键基因的重要新工具。
Expression of theFosBgene has been studied extensively in many fields using a variety of tools. However, previous techniques have had a variety of caveats, from potential off-target effects (e.g., overexpression of FosB, ΔFosB, or a dominant negative mutant of JunD, termed ΔJunD) or confounding developmental effects (e.g., the constitutiveFosBknockout mouse). Therefore, we sought to create afloxed FosBmouse line that will allow true silencing of theFosBgene with both spatial and temporal control. Here, we detail the cloning strategy, production, and validation of thefloxed FosBmouse. We demonstrate methodology for breeding and genotyping, and show that viral-mediated expression of Cre recombinase in a targeted, discrete brain region ablates expression of theFosBgene infloxedbut not wild type mice. Thus, thefloxed FosBmouse presented here represents an important new tool for the continued investigation of this critical gene.