Department of Chemical and Systems Biology, Stanford University, Stanford, California, USA.
PLoS One. 2013 Aug 22;8(8):e72393. doi: 10.1371/journal.pone.0072393. eCollection 2013.
Destabilizing domains are conditionally unstable protein domains that can be fused to a protein of interest resulting in degradation of the fusion protein in the absence of stabilizing ligand. These engineered protein domains enable rapid, reversible and dose-dependent control of protein expression levels in cultured cells and in vivo. To broaden the scope of this technology, we have engineered new destabilizing domains that perform well at temperatures of 20-25°C. This raises the possibility that our technology could be adapted for use at any temperature. We further show that these new destabilizing domains can be used to regulate protein concentrations in C. elegans. These data reinforce that DD can function in virtually any organism and temperature.
无定形结构域是条件不稳定的蛋白质结构域,可以与感兴趣的蛋白质融合,导致融合蛋白在没有稳定配体的情况下降解。这些工程化的蛋白质结构域能够在培养细胞和体内快速、可逆和剂量依赖性地控制蛋白质表达水平。为了扩大这项技术的应用范围,我们已经设计了新的在 20-25°C 温度下性能良好的去稳定结构域。这增加了我们的技术可以在任何温度下使用的可能性。我们还进一步表明,这些新的去稳定结构域可以用于调节秀丽隐杆线虫中的蛋白质浓度。这些数据进一步证明,DD 可以在几乎任何生物体和温度下发挥作用。