Efficient in vivo electroporation of the postnatal rodent forebrain.

Efficient in vivo electroporation of the postnatal rodent forebrain.
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DOI:
10.1371/journal.pone.0001883
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发表时间:
2008-04-02
期刊:
影响因子:
3.7
通讯作者:
Cremer H
Cremer H
中科院分区:
综合性期刊3区
文献类型:
--
作者:
Boutin C;Diestel S;Desoeuvre A;Tiveron MC;Cremer H

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体内功能基因分析仍然是生物医学研究的主要挑战。在这里,我们提出了一种新的方法,有效地引入核酸到出生后的小鼠前脑。我们发现,脑室内注射DNA,然后电穿孔诱导放射状胶质细胞,神经元前体细胞和嗅觉系统的神经元中的转基因的强表达。我们提出了两个证明的原理实验来验证我们的方法。首先,我们表明,在放射状神经胶质细胞中的神经细胞粘附分子(hNCAM-140)的人类同种型的表达诱导其分化成显示神经前体表型的细胞。第二,我们证明了p21作为一种细胞周期抑制剂,为出生后的神经干细胞。这种方法将代表一个重要的工具,为未来的研究出生后的神经发生和神经发育一般。
Functional gene analysis in vivo represents still a major challenge in biomedical research. Here we present a new method for the efficient introduction of nucleic acids into the postnatal mouse forebrain. We show that intraventricular injection of DNA followed by electroporation induces strong expression of transgenes in radial glia, neuronal precursors and neurons of the olfactory system. We present two proof-of-principle experiments to validate our approach. First, we show that expression of a human isoform of the neural cell adhesion molecule (hNCAM-140) in radial glia cells induces their differentiation into cells showing a neural precursor phenotype. Second, we demonstrate that p21 acts as a cell cycle inhibitor for postnatal neural stem cells. This approach will represent an important tool for future studies of postnatal neurogenesis and of neural development in general.
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期刊: SCIENCE
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