The invention relates to chemically inducible split protein actuators (CISPA), particularly to their design, manufacture, structure, and uses. The method of engineering CISPAs utilizes ligand- binding proteins or protein domains originating from humans or other organisms, which are rationally split into two fragments that reassemble only in the presence of a cognate ligand, which is typically a small molecule. The designed CISPAs can be used to regulate cellular processes such as gene expression, conditionally reconstitute of the function of a protein such as enzyme activity, as biological sensors, or for other applications.
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