Kidney-specific expression of GFP by in-utero delivery of pseudotyped adeno-associated virus 9.
Kidney-specific expression of GFP by in-utero delivery of pseudotyped adeno-associated virus 9.
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DOI：
10.1038/mtm.2014.14
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发表时间：
2014
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影响因子：
--
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以肾脏为靶点的基因治疗在很大程度上并不成功。最近，重组腺相关病毒(RAAV)载体被用于靶向成年小鼠肾脏。我们的假设是，伪型rAAV2/9载体可以在母鼠尾静脉注射怀孕小鼠后产生胎儿肾脏特异的绿色荧光蛋白(GFP)基因表达。用带有无处不在的巨细胞病毒启动子或最小的NPHS1启动子的rAAV2/9载体处理怀孕小鼠，以驱动GFP的肾脏特异性表达。对母鼠和幼崽的肾脏进行了载体DNA、基因表达和蛋白质的分析。12周后，在肾脏组织中发现了媒介DNA，水平较低但稳定，母鼠的水平高于幼鼠。在巨细胞病毒(CMV)增强的绿色荧光蛋白(EGFP)载体处理的母鼠和幼鼠的肾脏中都发现了GFP的强健表达。当用NPHS1-EGFP载体处理时，母鼠和幼鼠仅在肾脏中表达GFP，定位于肾小球。研究发现，到12周时，母犬的GFP mRNA表达增加了80倍，幼崽的GFP mRNA表达增加了近12倍。利用含有NPHS1启动子的伪型rAAV2/9载体，实现了基因治疗载体对胎肾的选择性靶向。
Gene therapy targeting of kidneys has been largely unsuccessful. Recently, a recombinant adeno-associated virus (rAAV) vector was used to target adult mouse kidneys. Our hypothesis is that a pseudotyped rAAV 2/9 vector can produce fetal kidney-specific expression of the green fluorescent protein (GFP) gene following maternal tail vein injection of pregnant mice. Pregnant mice were treated with rAAV2/9 vectors with either the ubiquitous cytomegalovirus promoter or the minimal NPHS1 promoter to drive kidney-specific expression of GFP. Kidneys from dams and pups were analyzed for vector DNA, gene expression, and protein. Vector DNA was identified in kidney tissue out to 12 weeks at low but stable levels, with levels higher in dams than that in pups. Robust GFP expression was identified in the kidneys of both dams and pups treated with the cytomegalovirus (CMV)-enhanced green fluorescent protein (eGFP) vector. When treated with the NPHS1-eGFP vector, dams and pups showed expression of GFP only in kidneys, localized to the glomeruli. An 80-fold increase in GFP mRNA expression in dams and a nearly 12-fold increase in pups was found out to 12 weeks of life. Selective targeting of the fetal kidney with a gene therapy vector was achieved by utilizing the pseudotyped rAAV 2/9 vector containing the NPHS1 promoter.
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影响因子：
12.4
作者：
Lipshutz, GS;Titre, D;Gaensler, KML
通讯作者：
Gaensler, KML
影响因子：
2.2
作者：
Lipshutz, GS;Flebbe-Rehwaldt, L;Gaensler, KNL
通讯作者：
Gaensler, KNL
影响因子：
6.1
作者：
Ahrabi, Ali K.;Jouret, Francois;Devuyst, Olivier
通讯作者：
Devuyst, Olivier
影响因子：
2
作者：
Martinovic-Bouriel, Jelena;Benachi, Alexandra;Gubler, Marie-Claire
通讯作者：
Gubler, Marie-Claire
DOI：
10.1006/bbrc.1999.0590
发表时间：
1999-05-10
影响因子：
3.1
作者：
Ochiya, T;Takahama, Y;Terada, M
通讯作者：
Terada, M