METHODOLOGICAL RECOMMENDATIONS FOR THE USE OF REPRODUCTIVE TECHNOLOGIES TO CREATE GENETICALLY MODIFIED RABBITS

Abstract

In this article, we present some practical recommendations that we developed in the course of two years of work on obtaining several lines of rabbits with an artificially modified genome. The technology of obtaining genetically modified rabbits, based on obtaining dated fertilized eggs with further microinjection of genetic constructs, assessment of survival and embryo transfer by laparoscopic and laparotomic methods. The following regimen demonstrated the greatest efficiency: administration of serum gonadotropin (Follimag, MOSAGROGEN, Russia) at a dose of 4ME/kg subcutaneously and intravenous administration of human chorionic gonadotropin (Chorulon, Merck Animal Health, USA) at a dose of 60ME/kg. Thus, the resulting operating procedure allows the operator to perform microinjections with high transfer efficiency and maximum cell survival. Microinjection is carried out through an Eppendorf FemtoJet 4i pneumatic microinjector using hydraulic micromanipulators under visual control through a NIKON ECLIPSE TS2R inverted research microscope. The described results may be useful in the compilation of standard operating procedures used in laboratories that are engaged in the production of genetically modified rabbit animals.