J Biol Chem, Vol. 273, Issue 14, 7928-7933, April 3, 1998
Receptor-mediated Transfection of Murine and Ovine Mammary Glands
in Vivo
Alexander S.
Sobolev
§,
Andrey A.
Rosenkranz§,
Olga
A.
Smirnova§,
Vladimir A.
Nikitin§,
Galina L.
Neugodova
,
Boris
S.
Naroditsky,
Ilya N.
Shilov§,
Ivan N.
Shatski§**, and
Lev K.
Ernst§
From the Department of Biophysics, Biological Faculty, Moscow
State University, 119889, the § Biophysical Laboratory and the
Laboratory of Genetic Engineering and Molecular Diagnostics of
Microorganisms, Russian Institute of Agricultural Biotechnology,
127550, Timiryazevskaya Street, 42, and the ** Belozersky Institute of
Physico-Chemical Biology, Moscow State University, 119899, Moscow,
Russia
Transfection of HC-11 murine epithelial mammary
cells as well as murine and sheep mammary glands were carried out using
insulin-containing constructs that deliver DNA by receptor-mediated
endocytosis to receptor-expressing cells. In vivo
transfection of mammary gland tissue with the luciferase gene was
carried out by introducing the DNA constructs into the mammary ducts of
both mice and sheep. The successful transfection of ewe mammary glands
was demonstrated by the detection of luciferase activity in mammary
gland biopsy material up to a month after a single administration of
the construct. To test whether products of expression of transfected
genes could be secreted into the milk in this system, the N-terminal
secretory signal sequences of bovine 
-lactoglobulin or the entire
coding sequence of human 
-lactalbumin were fused to the N terminus
of the luciferase gene. After transfection with the modified
luciferases, both murine and sheep milk could be shown to contain
luciferase activity, whereas mice, which had been transfected with the
nonmodified luciferase gene, did not secrete any activity in the milk.
This approach demonstrates for the first time the possibility of gene transfer in vivo into mammary gland epithelial cells using
constructs delivering DNA via receptor-mediated endocytosis.
Copyright © 1998 by The American Society for Biochemistry and Molecular Biology, Inc.
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