AbstractN,N‐dimethylaminobenzophenone (DMABP) represents an intramolecular donor–π–acceptor system resulting in luminescent properties that are highly sensitive to the local surrounding due to twisted intramolecular charge transfer (TICT). In this study, DMABP was covalently linked to a single (ss) or double‐stranded (ds) DNA and studied by a combination of UV/Vis absorption and fluorescence spectroscopy as well as femtosecond pump–probe measurements. As a result of embedding DMABP into a DNA environment, a drastic increase of the fluorescence quantum yield (QY) by a factor of more than 100 was detected, as further evidenced by an increase of the lifetime of the relevant excited states from less than 10 ps to more than 100 ps. A direct comparison between the ss‐ and ds‐DNA systems further demonstrates the high sensitivity to the surrounding area by means of a two‐fold difference in QY and fluorescence lifetimes. As a consequence, the respective DNA moiety (stacking and folding knot) affects significantly the competition between radiationless relaxation processes and fluorescence by interacting with DMABP.