title: Genomic integration of adenoviral gene transfer vectors following transduction of fertilized mouse oocytes.
abstract: Adenoviral vectors (AdV) are popular tools to deliver foreign genes into a wide range of cells. They have also been used in clinical gene therapy trials. Studies on AdV-mediated gene transfer to mammalian oocytes and transmission through the germ line have been reported controversially. In the present study we investigated whether AdV sequences integrate into the mouse genome by microinjecting AdV into the perivitelline space of fertilized oocytes. We applied a newly developed PCR technique (HiLo-PCR) for identification of chromosomal junctions next to the integrated AdV. We demonstrate that mouse oocytes can be transduced by different recombinant adenoviral vectors (first generation and gutless). In one transgenic mouse line using the first generation adenoviral vector, the genome has integrated into a highly repetitive cluster located on the Y chromosome. While the transgene (GFP) was expressed in early embryos, no expression was detected in adult transgenic mice. The use of gutless AdV resulted in expression of the transgene, albeit the vector was not transmitted to progeny. These results indicate that under optimized conditions fertilized mouse oocytes are transduced by AdV and give rise to transgenic founder animals. Therefore, adequate precautions should be taken in gene therapy protocols of reproductive patients since transduction of oocytes or early embryos and subsequent chromosomal integration cannot be ruled out entirely.
authors.affiliation: Department of Neurology Friedrich-Baur-Institute, Ludwig-Maximilians-Universität München, Munich, Germany. nancy.larochelle@mcgill.ca
authors.forename: Nancy
authors.initials: N
authors.lastname: Larochelle
authors.forename: Rolf
authors.initials: R
authors.lastname: Stucka
authors.forename: Norman
authors.initials: N
authors.lastname: Rieger
authors.forename: Lothar
authors.initials: L
authors.lastname: Schermelleh
authors.forename: Gudrun
authors.initials: G
authors.lastname: Schiedner
authors.forename: Stefan
authors.initials: S
authors.lastname: Kochanek
authors.forename: Eckhard
authors.initials: E
authors.lastname: Wolf
authors.forename: Hanns
authors.initials: H
authors.lastname: Lochmüller
mesh_terms: D000260:Adenoviruses, Human / Q000235:genetics*; D000818:Animals; D000971:Antineoplastic Combined Chemotherapy Protocols; D002945:Cisplatin; D004622:Embryo, Mammalian / Q000166:cytology / Q000821:virology*; D005260:Female; D018014:Gene Transfer Techniques; D005822:Genetic Vectors; D049452:Green Fluorescent Proteins / Q000235:genetics / Q000378:metabolism; D006801:Humans; D007069:Ifosfamide; D008297:Male; D051379:Mice; D008810:Mice, Inbred C57BL; D008811:Mice, Inbred DBA; D008822:Mice, Transgenic; D016685:Mitomycin; D009865:Oocytes / Q000821:virology*; D011995:Recombination, Genetic; D014161:Transduction, Genetic; D019076:Transgenes / Q000235:genetics / Q000502:physiology; D016662:Virus Integration
chemical_list: D049452:Green Fluorescent Proteins; D016685:Mitomycin; D002945:Cisplatin; D007069:Ifosfamide
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