L.A. Volkova, E.R. Mennibaeva, E.K. Tomgorova, A.N. Vetokh, N.A. Volkova, N.A. Zinovieva
L.K. Ernst All-Russian Research Institute of Animal Husbandry, pos. Dubrovitsy, 60, Podol’skii r-n, Moskovskaya obl., 142132, Russian Federation
Summary. Genetic modification of poultry by using lentiviral vectors is one of the most effective methods for producing transgenic animals with the desired properties. Considering physiological characteristics of birds, gene constructs can be introduced into target cells only at the multicellular embryo stage that causes the production of mosaic transgenic individuals. The aim of our study was to determine the optimal stage for the introduction of lentiviral vectors for transformation of chicken gonads in vivo. Studies were carried out on the basis of vivarium of the L.K. Ernst All-Russian Research Institute of Animal Husbandry in 2016. The transgenic poultry were obtained on the basis of Shaver Brown cross (n = 40) by using lentiviral vector pWCAG that contains the reporter gene eGFP (Green fluorescent protein) under control of a constitutive promoter CAG. The viral preparation was injected into chicken embryos at the 24th and 60th hours of incubation. The expression of eGFP in organs and tissues of transgenic chicken was determined by immunohistochemistry and fluorescence microscopy depending on the method of introducing the gene construct. A high percentage of the transformation of organs was observed with the injection of lentiviral vector into the chicken embryos at the 24th hour of incubation – (51.2 ± 6.3) %, which was on 6-8 % higher comparing to the introduction of the virus preparation at the 60th hour of incubation. However, the percentage of the transformation of reproductive organs was 1.9 times higher at the introduction of lentiviral vector in the dorsal aorta at the 60th hour of incubation. It may be due to the migration of primordial germ cells, which are precursors of germ cells, from blood in gonads at this stage. Thus, the 60th hour of incubation of embryos is the optimal period for introduction of lentiviral vectors into the chicken embryos in vivo to produce transgenic progeny.
Keywords: transgenesis, chickens, lentiviral vectors, expression, recombinant proteins.
Author Details: L.A. Volkova, Cand. Sc. (Biol.), leading research fellow (e-mail: Этот адрес электронной почты защищён от спам-ботов. У вас должен быть включен JavaScript для просмотра.); E.R. Mennibaeva, junior research fellow; E.K. Tomgorova, Cand. Sc. (Biol.), senior research fellow; A.N. Vetokh, junior research fellow; N.A. Volkova, D. Sc. (Biol.), head of laboratory; N.A. Zinovieva, D. Sc. (Biol.), director.
For citation: Volkova L.A., Mennibaeva E.R., Tomgorova E.K., Vetokh A.N., Volkova N.A., Zinovieva N.A. Efficiency of in Vivo Genetic Transformation of Chicken Gonads Depending on the Stage of Introduction of Lentiviral Vectors. Dostizheniya nauki i tekhniki APK. 2016. V. 30. No.11. Pp. 107-109 (in Russ.).