Dual 1 H and 19 F MR LacZ Gene Reporter Molecule
نویسندگان
چکیده
INTRODUCTION: The lacZ gene encoding enzyme beta-galactosidase (β-gal) is widely used as a reporter gene in diverse biological investigations including gene therapy. However, assessing β-gal activity in vivo remains challenging. Many colorimetric substrates have been developed over the years to assess β-gal activity and more recently in vivo imaging has been demonstrated based on fluorescence, bioluminescence, PET, SPECT, optoacoustics, and NMR. Notably, Meade et al. pioneered proton MRI contrast agents [1], and Mason et al. demonstrated F-labeled molecules as NMR active substrates [2]. Proton MRI alone can suffer from difficulty in identifying induced contrast in heterogeneous tissues, while F NMR often lacks SNR for effective imaging and is restricted to spectroscopy. A major strength of F NMR is the ability to observe substrate and product simultaneously based on chemical shift. A novel combined dual F/H MR lacZ gene detection approach based on (2-[(β-D-galactopyranosyl)oxy]3-fluorobenzaldehyde nicotinoyl hydrazone; GFBNHfigure below) was demonstrated in vitro, whereby F chemical shift reveals substrate and product and H MRI shows contrast from product [3]. We now show in vivo applications of GFBNH as a lacZ gene reporter molecule in mice.
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