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. With the advent of new endonuclease technology, single-stage mutation in several genes became possible, with the participation of one or both alleles (including obtaining homozygous animals in the F0 generation), introduction of several transgenes into the genome, and own gene replacement by a transgene. According to the demand for the use of new technologies, the production of animal biomodels with human genes for biomedical research is currently leading, but no less urgent task is to obtain animals that produce biologically active proteins with milk.
The purpose of this study is to create a genetic construct for the integration of the human lactoferrin cDNA transgene by the CRISPR/Cas HDR technology into the bovine beta-lactoglobulin locus (bβLg) with its simultaneous knockout on bovine microinjected zygotes. Based on the previously created plasmids β-LghLf and β-LghLfcmvEGFP, a genetic construct was obtained, including the cDNA sequences of hLF and the green fluorescent protein gene (EGFP) as a reporter, flanked by homology shoulders 5 ′ and 3 ’to the regulatory regions of bβLg. The design is supposed to be used to produce bovine embryos that are transgenic for hLf and/or βLgknockout.
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