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In this study, we report the rational design and synthesis of a novel 19F MRI probe that detects β-galactosidase (β-gal) activity, enabling the imaging of gene expression in cells. The 19F MRI signal of the probe was quenched by the intramolecular paramagnetic resonance enhancement from a Gd3+ ion. A contrivance was made in the probe structure to recover the 19F MRI signal after hydrolysis by β-gal with a following self-immolative reaction. This 19F MRI signal change was observed in the physiological aqueous condition. The probe could also detect β-gal activity in fixed HEK293T cells. In conclusion, this new probe enables the 19F MRI detection of cellular gene expression. The probe design strategy is also expected to lead to the development of MRI probes for a wide variety of hydrolase 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