A Protein-Tagging System for Signal Amplification in Gene Expression and Fluorescence Imaging

作者：Tanenbaum, Marvin E.; Gilbert, Luke A.; Qi, Lei S.; Weissman, Jonathan S.; Vale, Ronald D.^*

来源：Cell, 2014, 159(3): 635-646.

DOI：10.1016/j.cell.2014.09.039

• TRANSCRIPTION FACTOR SP1
• SYNERGISTIC ACTIVATION
• STEM-CELLS
• IN-VIVO
• RNA
• BINDING
• LOCALIZATION
• INTRABODIES
• HUNTINGTIN
• CRISPR/CAS

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.

• 出版日期2014-10-23