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Knock-out of genes in mature neurons by Cre-dependent CRISPR/Cas9 technique involving Cas9-cleaving sgRNAs to limit off-target effects.Delivery of miR-29a-3p to the arcuate hypothalamic nucleus attenuates obesity.This work significantly extends our understanding about the involvement of hypothalamic microRNAs in homeostatic regulation. Moreover, POMC Cre-dependent co-deletion of Nras in mature neurons attenuated miR-29 depletion-induced obesity. Within the latter, we identified a direct target of miR-29a-3p, Nras, which was up-regulated in those and only those mature POMC CreCas9 neurons that were effectively transduced by anti-miR-29 CRISPR-equipped construct.
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Moreover, we showed that miR-29 family is a prominent regulator of the PI3K-Akt-mTOR pathway. Here we found that mature neurons originating from this lineage employ miR-29a to protect against obesity, hyperphagia and insulin resistance, as demonstrated by POMC Cre-dependent CRISPR-Cas9 knock-out strategy in young or aged mice. This is achieved by various neurons many of which developmentally originate from the proopiomelanocortin (POMC)-expressing lineage. and Vinnikov I.A.✉Ībstract: Obesity, a growing threat to the modern society, represents an imbalance of metabolic queues that normally signal to the arcuate hypothalamic nucleus, a critic al brain region sensing and regulating energy homeostasis. Ma Y., Murgia N., Liu Y., Li Z., Sirakawin C., Konovalov R., Kovzel N., Xu Y., Kang X., Tiwari A., Mwangi P.M., Sun D., Erfle H., Konopka W., Lai Q., Najam S.S. Neuronal miR-29a protects from obesity in adult mice In perspective, the integration of expression and functional data from round worms, mouse and patient brain samples will provide novel insights for future genome wide association studies aiming to identify genetic variants within non-coding RNA genes and/or within the genes influenced by these RNA species. Our research group has a strong interest in studying the role of distinct neuronal population-specific coding and non-coding genes in chronic complex trait disorders and aging. Physiological and pathophysiological mechanisms involved in this regulation require intensive research to develop novel therapeutic strategies. They result from a failure in the interplay of environmental factors, humoral signals, extra- and intracellular enzymatic cascades, genetic and epigenetic signals in the central nervous system and in the periphe ry.
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