Baculoviral transduction of 3T3-L1 adipocytes with CRISPR/dCas9 system activating UCP-1 expression increases glucose uptakeстатьяЭлектронная публикацияТезисы
Статья опубликована в высокорейтинговом журнале
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Дата последнего поиска статьи во внешних источниках: 8 апреля 2022 г.
Аннотация:Nowadays beige‐like phenotype of adipocytes is considered to be a perspective therapeutic target to treat metabolic disorders, such as type 2 diabetes. Main function of these types of adipose tissue is to dissipate the proton gradient on mitochondria membrane via uncoupling protein 1 (UCP1) which leads to heat production. Activation of this protein expression in adipocytes could help obese people to dissipate extra energy and lower blood glucose level. Therefore, the purpose of this work was to investigate glucose uptake in cells that were transduced by baculovirus containing CRISPR/dCas9 system activating UCP‐1 expression. 3T3‐L1 preadipocytes were used as a beige adipocytes model. We carried out baculovirus transduction on 3T3‐L1 beige adipocytes according to a standard protocol. After this transduction we extracted RNA and performed reverse transcription reaction and real‐time PCR. After performing these steps we measured glucose uptake with [3H]‐deoxyglucose. We observed a significant increase in UCP‐1 gene expression in cells transduced with activating UCP‐1 system compaired to control group of non‐transduced cells and cells transduced with baculovirus without activating system. We showed that transduction with baculovirus decreased glucose uptake of 3T3‐L1 adipocytes compared to control, while glucose uptake of 3T3‐L1 adipocytes transduced with baculovirus containing CRISPR/dCas9 system activating UCP‐1 increased to the level of control group. Therefore, we can assume that activation of UCP‐1 increases glucose uptake due to the increase of cell energy needs and can be a possible therapeutic gene target to treat obesity.This work was supported by Russian Science Foundation grant #20‐45‐08003.