Tachikawa Kiyoshi, Schröder Oliver, Frey Gerhard, Briggs Steven P, Sera Takashi
Torrey Mesa Research Institute, 3115 Merryfield Row, San Diego, CA 92121, USA.
Proc Natl Acad Sci U S A. 2004 Oct 19;101(42):15225-30. doi: 10.1073/pnas.0406473101. Epub 2004 Oct 8.
We describe a facile method to activate or repress transcription of endogenous genes in a quantitative and specific manner by treatment with designed regulatory proteins (DRPs), in which artificial transcription factors (ATFs) are fused to cell-penetrating peptides (CPPs). Penetration of DRPs into cells is mediated by an N-terminal CPP fused to a nuclear localization signal; a DNA-binding domain and a transactivation domain follow. The DNA-binding domain was targeted to the vascular endothelial growth factor (VEGF)-A gene. An agonist DRP was rapidly taken up by cells and transported to the nucleus; soon after, the cells began transcribing the gene and secreting VEGF-A protein in a dose-dependent manner. Multiple copies of a short oligopeptide derived from a minimal transactivation domain of human beta-catenin was stronger than VP-16. The SRDX domain from the plant transcription factor, SUPERMAN, changed the DRP to a hypoxia-induced antagonist of VEGF-A. DRPs combine many of the potential benefits of transgenes with those of recombinant proteins.
我们描述了一种简便的方法，通过用设计的调节蛋白（DRP）处理，以定量和特异性的方式激活或抑制内源基因的转录，其中人工转录因子（ATF）与细胞穿透肽（CPP）融合。DRP进入细胞是由与核定位信号融合的N端CPP介导的；随后是DNA结合结构域和反式激活结构域。DNA结合结构域靶向血管内皮生长因子（VEGF）-A基因。一种激动剂DRP被细胞迅速摄取并转运到细胞核；不久之后，细胞开始转录该基因并以剂量依赖的方式分泌VEGF-A蛋白。源自人β-连环蛋白最小反式激活结构域的短寡肽的多个拷贝比VP-16更强。来自植物转录因子SUPERMAN的SRDX结构域将DRP转变为VEGF-A的缺氧诱导拮抗剂。DRP结合了转基因和重组蛋白的许多潜在益处。
