期刊
MOLECULAR THERAPY
卷 5, 期 5, 页码 501-508出版社
ACADEMIC PRESS INC ELSEVIER SCIENCE
DOI: 10.1006/mthe.2002.0577
关键词
gene transfer; in vivo delivery; naked DNA; ultrasound; in utero; oligodeoxynucleotide; microbubble; gene therapy
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.
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