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Receptor‐mediated transport of foreign DNA into preimplantation mammalian embryos
Author(s) -
Ivanova Margarita M.,
Rosenkranz Andrey A.,
Smirnova Olga A.,
Nikitin Vladimir A.,
Sobolev Alexander S.,
Landa Vladimir,
Naroditsky Boris S.,
Ernst Lev K.
Publication year - 1999
Publication title -
molecular reproduction and development
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.745
H-Index - 105
eISSN - 1098-2795
pISSN - 1040-452X
DOI - 10.1002/(sici)1098-2795(199910)54:2<112::aid-mrd2>3.0.co;2-u
Subject(s) - biology , zona pellucida , embryo , zygote , blastomere , microbiology and biotechnology , polylysine , naked dna , dna , oocyte , genetics , embryogenesis , biochemistry , plasmid
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. Mol. Reprod. Dev. 54:112–120, 1999. © 1999 Wiley‐Liss, Inc.