Systemic diffusion including germ cells after plasmidic in utero gene transfer in the rat

Fetal Diagnosis and Therapy
D GallotY J Bignon

Abstract

The integration of a proviral plasmid into the host genome could be a good approach for fetal somatic gene transfer. The goal of this study was to assess the integration and transcription of a proviral marker gene injected intraperitoneally into rat fetuses as well as the risks of maternal contamination and germ-line transmission. On day 17 post-coitus, each fetus was injected intraperitoneally with 10 microg plasmid DNA through the uterine wall. Twenty-one days after spontaneous delivery, integration and transcription of the plasmid in gonad, gut, liver, spleen, lung and brain tissue from 10 pups were determined by PCR and RT-PCR. 14 of 60 organs exhibited integration of the plasmid. Four samples of gut (40%), 3 samples of liver and spleen (30%), 2 samples of brain (20%) and no sample of lung were transfected. Two testicular samples were transfected and study of F1 rats from 2 brothers of one of the positive rats revealed transgenic pups from 1 of these 2 animals. No transfection of maternal tissues was detected. Integration and transcription of a marker gene injected intraperitoneally into rat fetuses appear efficient, especially in intraperitoneal organs. The risk of maternal contamination appears very low when using a naked...Continue Reading

Citations

Sep 8, 2005·Microbial Cell Factories·Jacob Glenting, Stephen Wessels
Apr 27, 2005·Molecular Therapy : the Journal of the American Society of Gene Therapy·Simon N WaddingtonJesus Prieto

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