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Receptor-mediated transport of foreign DNA into preimplantation mammalian embryos.
- Source :
-
Molecular reproduction and development [Mol Reprod Dev] 1999 Oct; Vol. 54 (2), pp. 112-20. - Publication Year :
- 1999
-
Abstract
- Mouse and rabbit preimplantation embryos with intact zona pellucida were incubated for 3 hr with DNA-carrying constructs containing insulin as an internalizable ligand: (insulin-polylysine)-DNA and (insulin-polylysine)-DNA-(streptavidin-polylysine)-(biotinylated adenovirus). Video-intensified microscopy demonstrated that the constructs penetrated the zona pellucida and accumulated in the blastomere perinuclear space. The percentage of blastocysts formed was about 70% after incubation of zygotes and two-cell embryos with the constructs. Foreign DNA was detected after 51 hr in 80% of rabbit embryos and after 96 hr in 73% of mouse embryos. Inclusion of various adenoviruses into the construct improved foreign DNA preservation in early embryos. Blot hybridization revealed genome-integrated foreign DNA in 12- and 15-day mouse embryos and in a newborn. Thus, the ligand-mediated mechanism can be employed for introducing foreign genetic material into early mammalian embryos; insulin provides for delivery inside the cell and to the nucleus, while adenoviruses ensure release from endosomes.<br /> (Copyright 1999 Wiley-Liss, Inc.)
- Subjects :
- Adenoviridae genetics
Animals
Blotting, Southern
Cells, Cultured
Embryo, Mammalian metabolism
Female
Insulin genetics
Insulin metabolism
Mice
Mice, Inbred BALB C
Microscopy, Video
Models, Genetic
Plasmids
Pregnancy
Rabbits
Transfection
DNA metabolism
Embryonic Development genetics
Endocytosis
Gene Transfer Techniques
Subjects
Details
- Language :
- English
- ISSN :
- 1040-452X
- Volume :
- 54
- Issue :
- 2
- Database :
- MEDLINE
- Journal :
- Molecular reproduction and development
- Publication Type :
- Academic Journal
- Accession number :
- 10471470
- Full Text :
- https://doi.org/10.1002/(SICI)1098-2795(199910)54:2<112::AID-MRD2>3.0.CO;2-U