Tri-part systems for protein dimerization and methods of use

Abstract

The present disclosure provides compositions and methods that utilize a target protein capable of binding to a small molecule to form a complex and a binding member that specifically binds to the complex wherein the target protein is derived from a non-human protein and the small molecule is an inhibitor of the non-human protein. The non-human protein may be derived from a viral, bacterial, fungal or protozoan protein. The compositions and methods allow for controlled interaction of polypeptides fused to the target protein and the binding member, respectively, and can be used to control the activity of dimerization inducible proteins, such as the activity of split transcription factors and split chimeric antigen receptors, by adding the small molecules. The disclosure provides expression vectors, binding members, dimerization inducible proteins, nucleic acids, cells, viral particles, kits, systems, and methods involving these components.

Description

[0001] This application claims priority to U.S. Provisional Application No. 62 / 874,025, filed July 15, 2019, the contents and elements of which are incorporated herein by reference for all purposes. technical field [0002] The present disclosure relates to compositions and methods that allow controlled interaction of polypeptides to which target proteins and binding members are fused. These compositions and methods utilize a target protein that binds to a small molecule forming a complex and a binding member that specifically binds the complex, wherein the target protein is derived from a non-human protein and the small molecule is a non-human protein. Non-human proteins can be derived from bacterial, viral, fungal or protozoan proteins. The non-human protein can be derived from a viral protease and the small molecule is a viral protease inhibitor. The present disclosure also relates to dimerization-inducible proteins comprising a target protein and a binding member, such as...

AI Technical Summary

Problems solved by technology

[0006] The approach described by Hill et al. 2018 and WO 2018 / 213848A1 to identify complex-specific molecules by utilizing existing small molecules and their targets is an attractive approach, however, certain human proteins (e.g. anti-apoptotic Bcl- xL protein) and the use of small molecules that bind to human targets in vivo are not without risk

For example, overexpression of a functional human protein can have effects on the cell expressing it, which may affect the health and viability of the cell

Furthermore, the use of small molecules whose targets are expressed in vivo results in increased dosage requirements due to binding competition of small molecules with endogenous and overexpressed targets

In addition, binding of small molecules to endogenous targets will affect the function of the protein, which may be detrimental to the cells expressing the target

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