Molotkov Dmitry A, Yukin Alexey Y, Afzalov Ramil A, Khiroug Leonard S
Neuroscience Center, University of Helsinki, Finland.
J Vis Exp. 2010 Sep 17(43):2244. doi: 10.3791/2244.
Creation of transgenic animals is a standard approach in studying functions of a gene of interest in vivo. However, many knockout or transgenic animals are not viable in those cases where the modified gene is expressed or deleted in the whole organism. Moreover, a variety of compensatory mechanisms often make it difficult to interpret the results. The compensatory effects can be alleviated by either timing the gene expression or limiting the amount of transfected cells. The method of postnatal non-ventricular microinjection and in vivo electroporation allows targeted delivery of genes, siRNA or dye molecules directly to a small region of interest in the newborn rodent brain. In contrast to conventional ventricular injection technique, this method allows transfection of non-migratory cell types. Animals transfected by means of the method described here can be used, for example, for two-photon in vivo imaging or in electrophysiological experiments on acute brain slices.
创建转基因动物是在体内研究感兴趣基因功能的一种标准方法。然而,在许多情况下,当修饰基因在整个生物体中表达或缺失时,许多基因敲除或转基因动物无法存活。此外,各种补偿机制常常使得难以解释结果。通过控制基因表达时间或限制转染细胞数量,可以减轻补偿效应。出生后非脑室显微注射和体内电穿孔方法允许将基因、小干扰RNA(siRNA)或染料分子直接靶向递送至新生啮齿动物大脑中的小目标区域。与传统的脑室注射技术相比,这种方法允许对非迁移性细胞类型进行转染。通过此处所述方法转染的动物可用于例如双光子体内成像或急性脑片的电生理实验。