Mandipropamid as a chemical inducer of proximity for in vivo applications

Abstract

Direct control of protein interactions by chemically induced protein proximity holds great potential for both cell and synthetic biology as well as therapeutic applications. Low toxicity, orthogonality and excellent cell permeability are important criteria for chemical inducers of proximity (CIPs), in particular for in vivo applications. Here, we present the use of the agrochemical mandipropamid (Mandi) as a highly efficient CIP in cell culture systems and living organisms. Mandi specifically induces complex formation between a sixfold mutant of the plant hormone receptor pyrabactin resistance 1 (PYR1) and abscisic acid insensitive (ABI). It is orthogonal to other plant hormone-based CIPs and rapamycin-based CIP systems. We demonstrate the applicability of the Mandi system for rapid and efficient protein translocation in mammalian cells and zebrafish embryos, protein network shuttling and manipulation of endogenous proteins.

A fluorogenic chemically induced dimerization technology for controlling, imaging and sensing protein proximity

“…CIP based on the rapamycin-inducible interaction of the FK506 binding protein (FKBP) and the FKBP-rapamycin binding domain (FRB) of the mammalian target of rapamycin (mTOR), is efficient and rapid, but is not reversible because of the high stability of the ternary assembly, and is limited by the biological activity of rapamycin 2,3 . These limits can be mitigated using non-toxic rapamycin analogs 4 or alternative technologies based on dimerization domains involved in the sensing of plant hormones [5][6][7][8] , although these latter can suffer from rather bulky dimerizing domains.…”

Engineered PROTAC-CID Systems for Mammalian Inducible Gene Regulation

“…For example, rapamycin is a widely used CID inducer but possesses undesirable immunosuppressive and autophagy-inducing effects. , Other CID inducers, such as abscisic acid (ABA) and gibberellic acid analogue (GA 3 ), are inefficient because they require high concentrations for ideal protein dimerization. Prior efforts to expand CID toolboxes include designing or mining new small molecules, − identifying new protein partners through screening nanobody/antibody libraries, , or computation-assisted protein design. , However, the number and type of highly efficient CIDs remain limited, preventing more sophisticated applications in mammalian cells or organisms.…”

Agrochemical control of gene expression using evolved split RNA polymerase

“…Mandipropamid and PYR1 MANDI protein constitute an engineered ligand-receptor pair, orthogonal to abscisic acid (ABA)-PYR1 receptor pair ( Park et al, 2015 ). The mandipropamid-PYR1 MANDI pair binds protein phosphatase HAB1 or ABI1 as the ABA-PYR1 pair ( Park et al, 2015 , Ziegler et al, 2022 and Fig. S1 ).…”