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.
journal_name
Mol Reprod Devjournal_title
Molecular reproduction and developmentauthors
Ivanova MM,Rosenkranz AA,Smirnova OA,Nikitin VA,Sobolev AS,Landa V,Naroditsky BS,Ernst LKdoi
10.1002/(SICI)1098-2795(199910)54:2<112::AID-MRD2>keywords:
subject
Has Abstractpub_date
1999-10-01 00:00:00pages
112-20issue
2eissn
1040-452Xissn
1098-2795pii
10.1002/(SICI)1098-2795(199910)54:2<112::AID-MRD2>journal_volume
54pub_type
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