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Expanding the Chemogenetic Toolbox by Circular Permutation.

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.

journal_name

J Mol Biol

journal_title

Journal of molecular biology

authors

Lee YT,He L,Zhou Y

doi

10.1016/j.jmb.2020.03.033

subject

Has Abstract

pub_date

2020-05-01 00:00:00

pages

3127-3136

issue

10

eissn

0022-2836

issn

1089-8638

pii

S0022-2836(20)30278-3

journal_volume

432

pub_type

杂志文章

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