1 Department of Molecular Neurobiology, Evotec AG , Hamburg, Germany .
Nucleic Acid Ther. 2013 Dec;23(6):389-400. doi: 10.1089/nat.2013.0445.
The repressor element-1 silencing transcription factor/neuron-restrictive silencer factor (REST/NRSF) is one of the first negative-acting transcriptional regulators implicated in vertebrate development thought to regulate hundreds of neuron-specific genes. However, its function in the adult system remains elusive. Here we employ second-generation antisense oligonucleotides (ASOs) to study the impact of rest-mediated suppression on gene expression. We demonstrate specific reductions in REST levels in vitro, and in vivo in mouse liver following treatment with ASOs, and we show that ASO mediated-REST suppression results in the elevation in expression of many neuronal genes including brain-derived neurotrophic factor, Synapsin1 (syn1) and β3-tubulin in BALB/c liver. Furthermore, we show the elevation of the affected proteins in plasma following ASO treatment. Finally, microarray analysis was applied to identify a broad range of genes modulated by REST suppression in mouse liver. Our findings suggest that REST may be an important target for neurodegenerative diseases like Huntington's disease, is also involved in the regulation of a broad range of additional cellular pathways, and that the antisense approach is a viable strategy for selectively modulating REST activity in vivo.
阻遏元件-1 沉默转录因子/神经元抑制因子（REST/NRSF）是第一个被认为参与脊椎动物发育的负调控转录因子之一，其被认为可以调控数百个神经元特异性基因。然而，其在成年系统中的功能仍不清楚。在这里，我们使用第二代反义寡核苷酸（ASO）来研究 REST 介导的抑制对基因表达的影响。我们证明了在体外和体内（在经过 ASO 处理的小鼠肝脏中），ASO 可以特异性地降低 REST 的水平，并且 ASO 介导的 REST 抑制导致包括脑源性神经营养因子、突触素 1（syn1）和β3-微管蛋白在内的许多神经元基因的表达升高。此外，我们还表明，在经过 ASO 处理后，受影响的蛋白质会在血浆中升高。最后，进行了微阵列分析以鉴定由 REST 抑制在小鼠肝脏中调节的广泛的基因。我们的研究结果表明，REST 可能是亨廷顿病等神经退行性疾病的一个重要靶点，它还参与了广泛的其他细胞途径的调节，并且反义方法是一种可行的策略，可以在体内选择性地调节 REST 活性。
