Transgenic rabbits with antisense RNA gene targeted at adenovirus H5
1991
Ernst, L.K. | Zakcharchenko, V.I. | Suraeva, N.M. | Ponomareva, T.I. | Miroshnichenko, O.I. | Prokof'ev, M.I. | Tikchonenko, T.I.
Production of transgenic rabbits by pronuclear microinjection of antisense RNA (asRNA) genes targeted against adenovirus h5 (Ad5) was performed. Efficacy of transgenosis, calculated as the ratio of transgenic to total born offspring, was equal to 36% in the case of linear plasmids. The copy number of integrated transgenes varied from 1 to 25 per genome. Only some of the integrated transgene copies cut by the appropriate restriction endonucleases yielded the hybridization bands identical to those of the original pGMA delta plasmid during blotting-analysis. The asRNA genes were retained in the offspring of transgenic does mated with wild-type bucks and were inherited at least in some transgenic animals in accordance to the normal Mendelian pattern. Transgenosis and crosses were accompanied by significant deletions and rearrangements of asRNA genes, so that their copy number varied independently among the members of the same brood. Nevertheless, at least two of five transgenic rabbits studied retained the intact asRNA genes. To evaluate the resistance to adenovirus driven by the asRNA genes, primary kidney cell cultures were raised from transgenic and normal rabbits and used for titration of Ad5. Two of four cell lines possessing the asRNA transgene were estimated to be 90% to 98% more resistant to Ad5 than a normal kidney cell line. Among seven primary kidney cell lines devoid of asRNA genes, only one manifested enhanced resistance to adenoviral infection.
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