Activation of UCP-1 expression in 3T3-L1 cells via CRISPR/dCas9 system leads to decrease of cytochrome c oxidase and superoxide dismutase genes expressionстатьяЭлектронная публикацияТезисы
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Дата последнего поиска статьи во внешних источниках: 8 апреля 2022 г.
Аннотация:Today obesity is one of the most widespread diseases among population. It is associated with white adipose tissue overgrowth. Brown‐like adipocytes called beige are differentiated from the same progenitor cells as white, but also are capable of producing heat. By switching white genotype to a beige, we could reprogram adipocytes to thermogenesis. Uncoupling protein 1 (UCP1) is the major player in this process; it decreases the proton gradient and produces heat. The aim of our work was to enhance UCP‐1 expression and investigate electron transport chain complexes' genes expression in 3T3‐L1 beige adipocytes after baculovirus transduction by CRISPR/dCas9 system for UCP‐1 gene activation. We differentiated 3T3‐L1 preadipocytes into beige adipocytes and performed baculovirus transduction via CRISPRa activating UCP‐1 according to standard protocols. Genes' expression was measured via Real‐time PCR. We have showed that indeed this activating system enhances UCP‐1 expression. Next thing we observed is a decrease of cytochrome c oxidase (Cox7) gene that can be explained by lowered activity of electron transport chain due to enhanced UCP‐1 expression. We also showed decrease of superoxide dismutase (SOD2) gene expression. We assume it can be explained by the fact that superoxide is a side product of cytochrome c oxidase activity, therefore when cytochrome c oxidase expression lowers the production of superoxide decreases. Thus in this work we have showed that enhanced UCP‐1 gene expression leads to decrease of cytochrome c oxidase and superoxide dismutase genes expression.This work was supported by Russian Science Foundation grant #20‐45‐08003.