Expanding the Chemogenetic Toolbox by Circular Permutation. Academic Article uri icon

abstract

  • To expand the repertoire of chemogenetic tools tailored for molecular and cellular engineering, we describe herein the design of cpRAPID as a circularly permuted rapamycin-inducible dimerization system composed of the canonical FK506-binding protein (FKBP) and circular permutants of FKBP12-rapamycin binding domain (cpFRB). By permuting the topology of the four helices within FRB, we have created cpFRB-FKBP pairs that respond to ligand with varying activation kinetics and dynamics. The cpRAPID system enables chemical-controllable subcellular redistribution of proteins, as well as inducible transcriptional activation when coupled with the CRISPR activation (CRISPRa) technology to induce a GFP reporter and endogenous gene expression. We have further demonstrated the use of cpRAPID to generate chemically switchable split nanobody (designated Chessbody) for ligand-gated antigen recognition in living cells. Collectively, the circular permutation approach offers a powerful means for diversifying the chemogenetics toolbox to benefit the burgeoning synthetic biology field.

published proceedings

  • J Mol Biol

altmetric score

  • 1.25

author list (cited authors)

  • Lee, Y., He, L., & Zhou, Y.

citation count

  • 2

complete list of authors

  • Lee, Yi-Tsang||He, Lian||Zhou, Yubin

publication date

  • May 2020