Nonviral gene transfer to surface skin of mid-gestational murine embryos by intraamniotic injection and subsequent electroporation

Molecular Reproduction and Development
Masahiro SatoNatsuko Kikuchi

Abstract

The surface epithelium of mid-gestational murine embryos is thought to be an attractive target for gene therapy in vivo, due to its visibility and accessibility from the external surface of the maternal uterus. Almost all studies of in utero gene transfer have adopted viral vectors for infection of fetal epithelium, and depended on intraamniotic introduction and simple incubation of vectors, leading to only infection of the surface layer (periderm) of fetal skin. Here we report a simple and convenient method of gene transfer of plasmid DNA into the deeper portion of surface skin of murine mid-gestational fetus. One to two microlitres of a solution containing a lacZ expression plasmid (0.5-1 microg) and trypan blue (0.05%) were placed onto the surface of a fetus (E 14.5) near the eye by a micropipette attached to a mouthpiece. This fetus was immediately electroporated by placing it between tweezer-type electrodes attached to a square-pulse generator. At 1 and 4 days after gene transfer, fetuses were subjected to histochemical staining for lacZ activity in the presence of X-Gal, a substrate for lacZ. Focal reactions were observed in the skin epidermal layers including periderm and basal layer 1 day after DNA introduction. However...Continue Reading

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Citations

Oct 21, 2015·Best Practice & Research. Clinical Obstetrics & Gynaecology·Lauren E McClain, Alan W Flake
Jun 30, 2009·Clinics in Perinatology·Matthew T SantoreAlan W Flake
May 28, 2011·Prenatal Diagnosis·Vedanta MehtaAnna L David
Jul 6, 2018·Archives of Pharmacal Research·Masahiro SatoChannabasavaiah B Gurumurthy

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