Controlled expression of transgenes introduced by in vivo electroporation
Controlled expression of transgenes introduced by in vivo electroporation
复制标题
DOI：
10.1073/pnas.0610155104
复制
发表时间：
2007-01-16
影响因子：
11.1
通讯作者：
Cepko, Constance L.
中科院分区：
文献类型：
--
作者：
Matsuda, Takahiko;Cepko, Constance L.
体内电穿孔是一种将基因导入生物体的强大技术。引入的基因或RNAi的表达的时间和空间调节将进一步增强该方法的实用性。在这里，我们证明了有条件的调控基因表达的电穿孔质粒在出生后的大鼠视网膜和胚胎小鼠大脑。对于时间调节，Cre/loxP介导的诱导型表达载体与表达Cre重组酶的条件活性形式的载体组合使用，所述Cre重组酶由4-羟基他莫昔芬激活。基因表达的起始受4-羟基他莫昔芬给药时间的调节。对于空间调节，通过使用对视杆光感受器、双极细胞、无长突细胞、Muller神经胶质或祖细胞特异性的启动子来表达转基因。这些构建体的组合将促进各种实验，包括细胞类型特异性基因错误表达、条件性RNAi以及祖细胞和前体细胞的命运作图。
In vivo electroporation is a powerful technique for the introduction of genes into organisms. Temporal and spatial regulation of expression of introduced genes, or of RNAi, would further enhance the utility of this method. Here we demonstrate conditional regulation of gene expression from electroporated plasmids in the postnatal rat retina and the embryonic mouse brain. For temporal regulation, Cre/loxP-mediated inducible expression vectors were used in combination with a vector expressing a conditionally active form of Cre recombinase, which is activated by 4-hydroxytamoxifen. Onset of gene expression was regulated by the timing of 4-hydroxytamoxifen administration. For spatial regulation, transgenes were expressed by using promoters specific for rod photoreceptors, bipolar cells, amacrine cells, Muller glia or progenitor cells. Combinations of these constructs will facilitate a variety of experiments, including cell-type-specific gene misexpression, conditional RNAi, and fate mapping of progenitor and precursor cells.