Всероссийский научно-исследовательский институт физиологии, биохимии и питания животных – филиал Федерального государственного бюджетного научного учреждения «Федеральный научный центр животноводства – ВИЖ имени академика Л.К. Эрнста»
ABSTRACT. The purpose of the article is to describe the main stages of the modified technology of producing transgenic mice, illustrated by the creating of humanized biomodels with integrated human N-acetyltransferase genes (Nat1 and Nat2). Improved method is described for inducing superovulation in mice-donors through the synchronization of sexual cycles in donor females at the time of their treatment by PMS gonadotrophin, which allows to get to a certain time 17 zygotes per donor with clearly visible pronuclei, suitable to the injecting of gene-engineered constructions. Linear fragments of the genetically engineered plasmid constructions comprising nucleotide sequences of human genes Nat1 Nat2 under mouse albumin gene promoter (hNat1i hNat2), were microinjected into the male pronuclei of zygotes derived from F1 hybrid female mice (CBA/la *C57BL/6). During preparation of the recipient females, the method was also used of preliminary replanting recipient females to vasectomized males through the hurdle, which helped to stimulate folliculogenesis and synchronization of sexual cycle in female recipient. So, it is possible to obtain a greater number of pseudopregnant female recipients with copulatory plugs to a certain time, which is one of the criteria of usefulness of the recipient animal, and the possibility of a successful transplantation of embryos microinjected with gene-engineered constructions. On the basis of these methods, a modified version of the technology for producing transgenic mice is designed. Transgenic mouses (F0) are produced with integrated into their genome constructions comprising the nucleotide sequences of human genes Nat1 Nat2 under mouse albumin gene promoter. Analysis of integration of transgenes in various tissues and organs in the offspring (F1) of the primary transgenic mice (F0) does indicate that the incorporation of human genes Nat1 and Nat2 happened to all three germ layers.
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