Fetal gene transfer by intrauterine injection with microbubble-enhanced ultrasound
Fetal gene transfer by intrauterine injection with microbubble-enhanced ultrasound
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DOI：
10.1006/mthe.2002.0577
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发表时间：
2002-05-01
影响因子：
12.4
通讯作者：
Kaneda, Y
中科院分区：
文献类型：
--
作者：
Endoh, M;Koibuchi, N;Kaneda, Y
宫内注射表达荧光素酶、绿色荧光蛋白（GFP）或β-半乳糖苷酶（β-gal）和异硫氰酸荧光素标记的寡脱氧核苷酸（FITC-ODN）的裸DNA，结合微泡增强超声（US），称为“鸟枪法”（SGM），在胎鼠中产生高水平的蛋白质表达。与单独注射裸DNA后的表达相比，使用SGM，荧光素酶表达增加了约10（3）倍。电子显微镜分析表明羊膜内（i.a.）注射SGM。在邻近注射点的多个胎儿组织中观察到GFP和β-gal的广泛表达以及FITC-ODN的递送。PCR分析表明，生殖细胞转染只是短暂的腹膜内（i.p）注射后，没有证据表明报告基因转移到后代。因此，SGM可能提供一个有用的手段，以澄清遗传性疾病的分子机制在子宫内，以及一个工具，开发基因治疗在子宫内。
Intrauterine injection of naked DNA expressing luciferase, green fluorescent protein (GFP), or beta-galactosidase (beta-gal) and fluorescein isothiocyanate-labeled oligodeoxynucleotide (FITC-ODN), in combination with microbubble-enhanced ultrasound (US), referred to as the "shot-gun method" (SGM), produced high-level protein expression in fetal mice. With the SGM, luciferase expression increased similar to10(3)-fold in comparison with expression after injection of naked DNA alone. Electron microscopic analysis demonstrated transient formation of pores on the skin surface after intraamniotic (i.a.) injection with the SGM. Widespread expression of GFP and beta-gal and delivery of FITC-ODN were observed in multiple fetal tissues adjacent to the injection points. PCR analysis indicated that germline transfection was only transient following intraperitoneal (i.p) injection, and there was no evidence of transfer of the reporter gene to the offspring. Thus, SGM might provide a useful means to clarify the molecular mechanisms of genetic diseases in utero, as well as a tool to develop gene therapies in utero.