A.N. Vetokh, N.A. Volkova, E.R. Mennibaeva, E.K. Tomgorova, I.K. Fomin, D.V. Beloglasov., 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. Vector systems based on the integrative viruses such as lentiviruses and retroviruses are an effective tool for the introduction and expression of genes. In this regard, the study of the conditions for transgene introduction into embryonic chicken cells is of interest, as well as the selection of regulatory sequences in the composition of gene constructs that provide high efficiency of transgenesis. The aim of the research was to study the influence of regulatory sequences in the composition of lentiviral vectors on development of chick embryos and effectiveness of obtaining chickens. The studies were carried out on the chicken embryo of Shaver Braun cross (n = 519) at the premises of vivarium of the L.K. Ernst Institute of Animal Husbandry in 2015-2016. pWpxl lentiviral vector was used in the study. Eight lentiviral vectors were obtained on its base, containing the reporter gene eGFP (green fluorescent protein) under the control of constitutive (pWCAG and pWRSV vectors) and tissue-specific (pW2.8, pW1.2, pWS1, pWS2, pWM1 and pWM2 vectors) regulatory elements. Viral preparation was injected into chicken embryos at the 24th and 60th hours of incubation. Dynamics of embryo development and efficiency of poultry obtaining were studied depending on introduction method for lentiviral vectors and regulatory sequences in their composition. The expression of the recombinant protein in the embryonic cells was determined at the sixth day of incubation using flow cytofluorometry. A relatively high percentage of embryonic cells that expressed recombinant protein eGFP were established at the transfection of embryonic cells at the 24th hour of incubation – 34.3 %, which was higher by 4.3 % compared with the administration of lentiviral vectors in the dorsal aorta of embryos at the 60th hours of incubation. The least effect on the embryogenesis was observed using lentiviral vectors with regulatory elements that provide tissue specific expression of recombinant genes. The percentage of embryos that developed, when lentiviral vectors with a constitutive expression of the structural gene was used, was lower on average by 6.5 %.
Keywords: transgenesis, chickens, lentiviral vectors, expression, recombinant proteins.
Author Details: A.N. Vetokh, junior research fellow (e-mail Этот адрес электронной почты защищён от спам-ботов. У вас должен быть включен JavaScript для просмотра.); N.A. Volkova, D. Sc. (Biol.), head of laboratory; E.R. Mennibaeva, junior research fellow; E.K. Tomgorova, Cand. Sc. (Biol.), senior research fellow; I.K. Fomin, Cand. Sc. (Biol.), senior research fellow; D.V. Beloglasov., Cand. Sc. (Biol.), research fellow; N.A. Zinovieva, D. Sc. (Biol.), member of the RAS, director.
For citation: Vetokh A.N., Volkova N.A., Mennibaeva E.R., Tomgorova E.K., Fomin I.K., Beloglasov D.V., Zinovieva N.A. Effectiveness of Chick Embryo Development with the Introduction of Lentiviral Vectors in Vivo. Dostizheniya nauki i tekhniki APK. 2016. V. 30. No 12. Pp. 83-85 (in Russ.).