Biopolym. Cell. 1990; 6(2):17-24.
The transgenic rabbits containing a human apoA-i antisense gene as a model for genetic correction of atherogenic disturbances of lipid metabolism
1Perevozchikov A. P., 1Valsman B. L., 1Dozortsev D. I., 1Sorokin A. V., 1Orlov S. V., 1Denisenko A. D., 1Dyban A. P., 1Klimov A. N.
  1. Institute of Experimental Medicine, Academy of Medical Sciences of the USSR
    Leningrad, USSR

Abstract

The recombinant DNA molecules containing potentially expressing human apoA-I antisence gene were constructed and injected into the rabbit zygotes. Dot-blot hybridization techniques as well as lipid and lipoprotein analyses methods were used to reveal transgenic animals. It was found that DNAs of several rabbits contained up to 40 copies of molecules of foreign DNA per genome. It was interesting that the patterns of serum lipid and lipoprotein metabolism of transgenic animals were shifted to atherogenic phenotype. In one case the expression of the antisense gene was able to suppress the synthesis of high-density lipoproteins. In two other cases the expression of this gene as appeared was correlated with a decrease of α-lipoprotein cholesterol level. In all the cases serum cholesterol and (or) triglyceride levels increased as well. Thus, the obtained transgenic rabbits may serve as a model to study and genetically correct atherogenic disturbances of lipid metabolism.

References

[1] Klimov AI. Causes and conditions for the development of atherosclerosis. Biochemical basis of the pathogenesis of atherosclerosis. Leningrad, 1980; 3-45.
[2] Wilson JM, Johnston DE, Jefferson DM, Mulligan RC. Correction of the genetic defect in hepatocytes from the Watanabe heritable hyperlipidemic rabbit. Proc Natl Acad Sci U S A. 1988;85(12):4421-5.
[3] Miyanohara A, Sharkey MF, Witztum JL, Steinberg D, Friedmann T. Efficient expression of retroviral vector-transduced human low density lipoprotein (LDL) receptor in LDL receptor-deficient rabbit fibroblasts in vitro. Proc Natl Acad Sci U S A. 1988;85(17):6538-42.
[4] van der Krol AR, Mol JN, Stuitje AR. Modulation of eukaryotic gene expression by complementary RNA or DNA sequences. Biotechniques. 1988;6(10):958-76. Review.
[5] Dudov KP, Perry RP. The gene family encoding the mouse ribosomal protein L32 contains a uniquely expressed intron-containing gene and an unmutated processed gene. Cell. 1984;37(2):457-68.
[6] Cheung P, Chan L. Nucleotide sequence of cloned cDNA of human apolipoprotein A-I. Nucleic Acids Res. 1983;11(11):3703-15.
[7] Vaysman BL, Golinskiy GF. Technique of microinjection of cloned DNA fragments (foreign genes) into mouse feminonucleus. General patterns and control mechanisms of early embryogenesis of mammals in normal and pathological. Ed. AP Dyban. Leningrad, Medicine, 1985; 108-13.
[8] Magracheva EIa. Separation of lipoproteins from blood serum by polyacrylamide disc electrophoresis. Vopr Med Khim. 1973;19(6):652-5.
[9] Dudov KP, Perry RP. Properties of a mouse ribosomal protein promoter. Proc Natl Acad Sci U S A. 1986;83(22):8545-9.
[10] Borresen AL. High density lipoprotein (HDL) polymorphisms in rabbit. I. A comparative study of rabbit and human serum high density lipoprotein. J Immunogenet. 1976;3(2):73-81.
[11] Haddad IA, Ordovas JM, Fitzpatrick T, Karathanasis SK. Linkage, evolution, and expression of the rat apolipoprotein A-I, C-III, and A-IV genes. J Biol Chem. 1986;261(28):13268-77.
[12] Klimov AN, Nikul'cheva NG. Lipoproteides, dyslipoproteinemia and atherosclerosis. Leningrad, Medicine, 1984; 223 p.
[13] Va?sman BL, Kapelinskaia TV, Gorodetski? SI, Dyban AP. Possibility of breeding of animals-producers of biologically active substances--by microinjection of cloned genes into the ovum. Antibiot Khimioter. 1988;33(2):154-8.
[14] Karathanasis SK. Apolipoprotein multigene family: tandem organization of human apolipoprotein AI, CIII, and AIV genes. Proc Natl Acad Sci U S A. 1985;82(19):6374-8.