The efficiency of use lentiviral vectors for the transformation of rooster spermatogenic cells in vivo

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Abstract


Male gonad cells are considered as promising target cells for the introduction of recombinant DNA within obtaining genetically modified individuals with given characteristics. The use of testicular spermatogonial stem cells is of the greatest interest. In the process of differentiation, this type of cell gives rise to a significant population of mature male germ cells. In the case of their genetic transformation, differentiated cells can be used to inseminate females in order to produce transgenic progeny. The aim of the research was to study the efficiency of using lentiviral vectors for the local transformation of roosters’ testicular spermatogenic cells. We used a lentiviral vector containing the ZsGreen reporter gene under the control of the CMV promoter. In vitro transformation of rooster spermatogenic cells was carried out by infection with a viral preparation, in vivo through multiple injections of the viral preparation into the testicular parenchyma of roosters ( n = 5). The efficiency of transformation was assessed by expression of the reporter ZsGreen gene in transfected spermatogenic cells. The success of using lentiviral vectors for the genetic transformation of rooster spermatogenic cells was shown in experiments in vitro and in vivo . The transformation efficiency of this cells types in an in vitro culture varied from 45 to 57% and averaged 48 ± 4%. The expression of the ZsGreen reporter gene in the cells of the spermatogenic epithelium of the testes was established in almost all experimental roosters in the in vivo experiments. The number of seminiferous tubules with transformed spermatogenic cells varied in the studied experimental roosters from 10 to 22%. The effectiveness of genetic transformation of the testes spermatogenic cells was 1.8 ± 0.2%. The obtained results indicate to the success of using lentiviral vectors for the genetic transformation of spermatogenic cells of rooster testes in vivo in order to create individuals with genetically transformed germ cells for the further production of transgenic offspring with given characteristics.


About the authors

Natalya Alexandrovna Volkova

Federal Science Center for Animal Husbandry named after Academy Member L.K. Ernst

Author for correspondence.
Email: natavolkova@inbox.ru
Moscow region, Russian Federation

Doctor of Sciences in Biology, Professor of the Russian Academy of Sciences, Head of the Laboratory of Cell Engineering

Anastasia Nikolaevna Vetokh

Federal Science Center for Animal Husbandry named after Academy Member L.K. Ernst

Email: anastezuya@mail.ru
Moscow region, Russian Federation

Researcher of the Laboratory of Functional and Evolutionary Genomics of Animals

Lyudmila Aleksandrovna Volkova

Federal Science Center for Animal Husbandry named after Academy Member L.K. Ernst

Email: ludavolkova@inbox.ru
Moscow region, Russian Federation

Candidate of Sciences in Biology, Senior Researcher of the Laboratory of Cell Engineering

Anatolievna Zinovyeva Nataliya

Federal Science Center for Animal Husbandry named after Academy Member L.K. Ernst

Email: n_zinovieva@mail.ru
Moscow region, Russian Federation

Doctor of Sciences in Biology, Academician of the Russian Academy of Sciences

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Copyright (c) 2019 Volkova N.A., Vetokh A.N., Volkova L.A., Nataliya A.Z.

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