title: Novel NMR platform for detecting gene transfection: synthesis and evaluation of fluorinated phenyl beta-D-galactosides with potential application for assessing LacZ gene expression.
abstract: Gene therapy holds great promise for the treatment of diverse diseases, but widespread implementation is hindered by difficulties in assessing the success of transfection. The development of noninvasive reporter techniques based on appropriate molecules and imaging modalities may help to assay gene expression. Fluorophenyl-beta-d-galactopyranosides provide a novel class of NMR active molecules, which are highly responsive to the action of beta-galactosidase (beta-gal), the product of the lacZ gene. The reporter molecules are stable in solution and with respect to wild-type cells, but the enzyme causes liberation of the aglycon, a fluorophenol, accompanied by distinct color formation and a (19)F NMR chemical shift of 5-10 ppm, depending on pH. Synthetic strategy, experimental methods, and molecular and (19)F NMR characteristics are reported for a series of molecules in solution, blood, and tumor cells. This class of molecules presents a new strategy for assaying gene expression with a highly versatile molecular structural platform.
authors.affiliation: Department Radiology, The University of Texas Southwestern Medical Center at Dallas, Dallas, Texas 75390-9058, USA.
authors.forename: Jianxin
authors.initials: J
authors.lastname: Yu
authors.forename: Pieter
authors.initials: P
authors.lastname: Otten
authors.forename: Zhenyi
authors.initials: Z
authors.lastname: Ma
authors.forename: Weina
authors.initials: W
authors.lastname: Cui
authors.forename: Li
authors.initials: L
authors.lastname: Liu
authors.forename: Ralph P
authors.initials: RP
authors.lastname: Mason
mesh_terms: D000818:Animals; D045744:Cell Line, Tumor; D004353:Drug Evaluation, Preclinical / Q000379:methods; D005461:Fluorine / Q000032:analysis / Q000737:chemistry*; D005697:Galactosides / Q000032:analysis / Q000138:chemical synthesis*; D015972:Gene Expression Regulation, Neoplastic; D007763:Lac Operon; D019906:Nuclear Magnetic Resonance, Biomolecular / Q000379:methods*; D051381:Rats; D014162:Transfection / Q000379:methods*
chemical_list: D005697:Galactosides; C007816:beta-galactoside; D005461:Fluorine