Abstract: Provided are methods of assessing activity of a promoter region. The methods include culturing a cell including a nucleic acid, the nucleic acid including a region that encodes an enzyme donor (ED) operably coupled to a promoter region, under conditions in which the ED is expressed when the promoter region is active. The methods further include contacting the ED, if expressed, with an enzyme acceptor (EA) to form ED-EA complexes having enzymatic activity. The methods further include detecting the level of the enzymatic activity to assess activity of the promoter region. Activity of the promoter region may be indicative, and therefore may be used to assess, the activity of a cellular signaling pathway of interest and/or of endogenous or exogenous (e.g., introduced) transcription factors of interest. Cells, compositions, and kits that find use, e.g., in practicing the methods of the present disclosure, are also provided.