Lentiviral transduction of neuronal cells.
Lentiviral transduction of neuronal cells.
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DOI：
10.1007/978-1-62703-640-5_12
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发表时间：
2013
期刊：
影响因子：
--
通讯作者：
White, Martyn K
中科院分区：
文献类型：
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作者：
Wollebo, Hassen S;Woldemichaele, Baheru;White, Martyn K
在这里，我们描述了一种构建慢病毒载体的一般方法，使用一个特定的例子，在两个假设的启动子和衍生的基于HIV-1的慢病毒表达载体pLVX-Puro的控制下构建含有荧光素酶报告基因的慢病毒。该方法可用于比较不同启动子的强度和调控。在本例中，转导的靶细胞是人原代胎儿星形胶质细胞，但该方法适用于来自中枢神经系统或其他组织的任何原代细胞培养，并可用于检查不同细胞类型中特定启动子的强度。用Fugene 6转染试剂将4个质粒转染到293T细胞中制备HIV慢病毒颗粒。
Here we describe a general method for the construction of a lentivirus vector using a specific example of the construction of a lentivirus containing the luciferase reporter gene under the control of two hypothetical promoters and derived HIV-1 based lentivirus expression vector pLVX-Puro. This method can be used to compare the strength and regulation of different promoters. In this example, the target cells for transduction are human primary fetal astrocytes but the method is applicable to any primary cell culture from the CNS or other tissue and can be used to examine the strength of a particular promoter in different cell types. HIV based lentivirus particles are prepared by transfection of 4 plasmids into 293T cells using the Fugene 6 transfection reagent.