Tanenbaum Marvin E, Gilbert Luke A, Qi Lei S, Weissman Jonathan S, Vale Ronald D
Department of Cellular and Molecular Pharmacology, University of California, San Francisco, San Francisco, CA 94158, USA; Howard Hughes Medical Institute, University of California, San Francisco, San Francisco, CA 94158, USA.
Department of Cellular and Molecular Pharmacology, University of California, San Francisco, San Francisco, CA 94158, USA; Howard Hughes Medical Institute, University of California, San Francisco, San Francisco, CA 94158, USA; Center for RNA Systems Biology, University of California, Berkeley, Berkeley, CA 94720, USA; California Institute for Quantitative Biomedical Research (QB3), San Francisco, CA 94158, USA.
Cell. 2014 Oct 23;159(3):635-46. doi: 10.1016/j.cell.2014.09.039. Epub 2014 Oct 9.
Signals in many biological processes can be amplified by recruiting multiple copies of regulatory proteins to a site of action. Harnessing this principle, we have developed a protein scaffold, a repeating peptide array termed SunTag, which can recruit multiple copies of an antibody-fusion protein. We show that the SunTag can recruit up to 24 copies of GFP, thereby enabling long-term imaging of single protein molecules in living cells. We also use the SunTag to create a potent synthetic transcription factor by recruiting multiple copies of a transcriptional activation domain to a nuclease-deficient CRISPR/Cas9 protein and demonstrate strong activation of endogenous gene expression and re-engineered cell behavior with this system. Thus, the SunTag provides a versatile platform for multimerizing proteins on a target protein scaffold and is likely to have many applications in imaging and controlling biological outputs.
在许多生物过程中，信号可以通过招募多个调控蛋白拷贝至作用位点来实现放大。利用这一原理，我们开发了一种蛋白质支架，即一种名为SunTag的重复肽阵列，它可以招募多个抗体融合蛋白拷贝。我们证明，SunTag可以招募多达24个绿色荧光蛋白（GFP）拷贝，从而能够对活细胞中的单个蛋白质分子进行长期成像。我们还利用SunTag，通过将转录激活结构域的多个拷贝招募至核酸酶缺陷型CRISPR/Cas9蛋白，创建了一种高效的合成转录因子，并证明该系统能强烈激活内源性基因表达并重塑细胞行为。因此，SunTag为在目标蛋白质支架上使蛋白质多聚化提供了一个通用平台，并且可能在成像和控制生物学输出方面有许多应用。
