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Cre-mediated transgene activation in the developing and adult mouse brain.

机译:Cre介导的转基因激活在发育中和成年小鼠的大脑中。

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The neuron-specific rat enolase (NSE) promoter was employed to establish transgenic mice expressing Cre recombinase in the central nervous system. Founders were crossed with dormant lacZ indicator mice and specificity as well as efficiency of Cre-mediated transgene activation was determined by PCR and/or X-gal staining. Whereas most transgenic lines exhibited Cre activity in early development resulting in widespread Cre activity, one line (NSE-Cre26) expressed high levels of Cre in the developing and adult brain. With the exception of kidney, which showed occasionally low level of Cre activity, Cre recombination in double transgenics was restricted to the nervous system. Whole-mount X-gal staining of 9.5 dpc embryos indicated Cre-mediated lacZ expression in forebrain, hindbrain, and along the midbrain flexure. A similar expression pattern was observed during later stages of embryogenesis (11.5-13.5 dpc). In adult mice, Cre recombinase was expressed in cerebral cortex and cerebellum and high levels of Cre-mediated lacZ expression were observed in hippocampus, cortex, and septum. The NSE-Cre26 transgenic mouse line thus provides a useful tool to specifically overexpress and/or inactivate genes in the developing and adult brain.
机译:使用神经元特异性大鼠烯醇化酶(NSE)启动子来建立在中枢神经系统中表达Cre重组酶的转基因小鼠。创始人与休眠的lacZ指标小鼠杂交,并通过PCR和/或X-gal染色确定Cre介导的转基因激活的特异性和效率。大多数转基因品系在早期发育中表现出Cre活性,从而导致广泛的Cre活性,而一个品系(NSE-Cre26)在发育中和成年大脑中表达高水平的Cre。除了肾脏偶尔显示出较低的Cre活性水平外,双转基因中的Cre重组仅限于神经系统。 9.5 dpc胚胎的X-gal完整染色表明,Cre介导的lacZ在前脑,后脑和中脑弯曲处表达。在胚胎发生的后期(11.5-13.5 dpc)观察到了相似的表达模式。在成年小鼠中,Cre重组酶在大脑皮层和小脑中表达,在海马,皮层和中隔中观察到高水平的Cre介导的lacZ表达。因此,NSE-Cre26转基因小鼠品系提供了一种有用的工具,可以在发育中的和成年的大脑中特异性地过表达和/或灭活基因。

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