Adenovirus-mediated gene transfer in the midgestation fetal mouse
Adenovirus-mediated gene transfer in the midgestation fetal mouse
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DOI：
10.1006/jsre.1999.5588
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发表时间：
1999-06-15
影响因子：
2.2
通讯作者：
Gaensler, KNL
中科院分区：
文献类型：
--
作者：
Lipshutz, GS;Flebbe-Rehwaldt, L;Gaensler, KNL
背景子宫内基因转移战略的发展将使改善并最终治愈与产前和产后发病率和死亡率有关的遗传疾病成为可能。我们已经开发了一种小鼠模型，在子宫内，肝内，腺病毒介导的基因转移在第15天的胎儿，并比较了荧光素酶报告基因表达的水平和分布在新生儿与成年动物静脉注射。CD-1胎仔在妊娠第15天肝内注射1 × 107颗粒形成单位（PFU）的E1和E3缺失重组腺病毒（含有荧光素酶报告基因）或生理盐水。出生后，将幼鼠安乐死，收获脑、心、肠、肝、肺和脾并分析荧光素酶活性。两个腺病毒注射窝进行足月和一个雌性流产。分析了实验组中10只新生小鼠和对照组中5只新生小鼠的组织;保留其余新生小鼠的组织用于其他研究。在所有腺病毒注射的新生儿肝脏中检测到高水平的荧光素酶表达。在远处器官中检测到较低水平的荧光素酶活性。通过血清转氨酶升高确定的肝毒性在成年小鼠中观察到，但在先前注射腺荧光素酶病毒的新生小鼠中未观察到。在子宫内肝内基因传递与腺病毒载体在发育中的小鼠胎儿是可行的，并产生高水平的基因表达。这些研究表明，病毒和非病毒的基因传递载体可能是有用的，在未来的方法来产前治疗遗传性疾病的发展。(C)北京：科学出版社.
Background. The development of strategies for gene transfer in utero will make possible the amelioration, and eventually the cure, of genetic diseases associated with pre- and postnatal morbidity and mortality. We have developed a murine model for in utero, intrahepatic, adenovirus-mediated gene transfer in Day 15 fetuses and compared the level and distribution of luciferase reporter gene expression in newborns with those observed in adult animals injected intravenously.Material and methods. CD-1 fetuses underwent intrahepatic injection on Day 15 of gestation with 1 x 10(7) particle-forming units (PFU) of an E1- and E3-deleted recombinant adenovirus containing the luciferase reporter gene or with normal saline. At birth, pups were euthanized, and the brain, heart, intestine, liver, lungs, and spleen harvested and analyzed for luciferase activity.Results. Two adenovirus-injected litters proceeded to term and one female aborted. Tissues from 10 newborn mice in the experimental group and 5 newborns in the control group were analyzed; tissues from the remaining newborns were reserved for other studies. High-level luciferase expression was detected in all adenovirus-injected newborn livers. Lower levels of luciferase activity were detected in distant organs. Hepatic toxicity as determined by serum transaminase elevations was observed in adult, but not in newborn mice previously injected with the adeno-luciferase virus.Conclusions. In utero intrahepatic gene delivery with adenoviral vectors in the developing murine fetus is feasible and produces high-level gene expression. These studies suggest that viral and nonviral gene delivery vectors may be useful in the development of future approaches to prenatal treatment of genetic disorders. (C) 1999 Academic Press.