Frosted DNA: β‐Galactosidase Control of Oligonucleotide Activity

• DOI: 10.1002/chem.202500347
• Publication Date: 2025-03-14T08:45:29Z
• Authors: Fabian Sinsel; Marius Külp; Michael A. Rieger; Alexander Heckel

ABSTRACTWe introduce a novel approach for the control of oligonucleotides through enzymatic activation with β‐galactosidase (β‐gal). We use the well‐known enzymatic capability of β‐gal to hydrolyze the β‐galactosidic bond combined with a self‐immolative linker and present three ways of steric or topological blocking of a DNA oligonucleotide. Through a series of in vitro experiments with β‐gal variants (recombinant or from human cell lysates), we systematically investigate stability, transitory perturbation, and enzymatic activation. Our approach holds significant promise for applications related to senescence‐associated β‐gal activity, including targeted modulation of gene expression and programmable molecular interventions. The combination of enzymatic activation applied to oligonucleotides represents a significant advance for targeted release in affected cells without the need for external triggering.