Production of a Cloned Offspring and CRISPR/Cas9 Genome Editing of Embryonic Fibroblasts in Cattle.

Autor: Singina GN; Ernst Federal Science Center for Animal Husbandry, Podolsk, Russia. g_singina@mail.ru., Sergiev PV; Institute of Functional Genomics, Moscow State University, Moscow, Russia.; Center of Life Sciences, Skolkovo Institute of Science and Technology, Skolkovo, Russia.; Faculty of Chemistry, Moscow State University, Moscow, Russia., Lopukhov AV; Ernst Federal Science Center for Animal Husbandry, Podolsk, Russia., Rubtsova MP; Faculty of Chemistry, Moscow State University, Moscow, Russia., Taradajnic NP; Ernst Federal Science Center for Animal Husbandry, Podolsk, Russia., Ravin NV; Research Center of Biotechnology, Moscow, Russia., Shedova EN; Ernst Federal Science Center for Animal Husbandry, Podolsk, Russia., Taradajnic TE; Ernst Federal Science Center for Animal Husbandry, Podolsk, Russia., Polejaeva IA; Department of Animal, Dairy and Veterinary Sciences, Utah State University, Logan, UT, USA., Dozev AV; Ernst Federal Science Center for Animal Husbandry, Podolsk, Russia., Brem G; Department of Animal Breeding and Genetics, University of Veterinary Medicine, Vienna, Austria., Dontsova OA; Center of Life Sciences, Skolkovo Institute of Science and Technology, Skolkovo, Russia.; Faculty of Chemistry, Moscow State University, Moscow, Russia.; Belozersky Research Institute of Physico-Chemical Biology, Moscow State University, Moscow, Russia.; Shemyakin-Ovchinnikov Institute of Bioorganic Chemistry, Moscow, Russia., Zinovieva NA; Ernst Federal Science Center for Animal Husbandry, Podolsk, Russia.
Jazyk: angličtina
Zdroj: Doklady. Biochemistry and biophysics [Dokl Biochem Biophys] 2021 May; Vol. 496 (1), pp. 48-51. Date of Electronic Publication: 2021 Mar 10.
DOI: 10.1134/S1607672921010099
Abstrakt: Somatic Cell Nuclear Transfer (SCNT) technique was used to produce the first viable cloned cattle offspring in Russia. Whole-genome SNP genotyping confirmed that the cloned calf was identical to the fibroblast cell line that was used for SCNT. CRISPR/Cas9 approach was subsequently used to knock out genes for beta-lactoglobulin gene (PAEP) and the beta-lactoglobulin-like protein gene (LOC100848610) in the fibroblast cells. Gene editing (GE) efficiency was 4.4% for each of these genes. We successfully obtained single-cell-derived fibroblast colonies containing PAEP and LOC100848610 knockouts, which will be used to produce beta-lactoglobulin-deficient cattle.
Databáze: MEDLINE
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