Synthesis and evaluation of novel enhanced gene reporter molecules: detection of beta-galactosidase activity using 19F NMR of trifluoromethylated aryl beta-D-galactopyranosides.
Gene therapy has emerged as a promising strategy for treatment of various diseases, but there is a pressing need for the development of non-invasive reporter techniques based on appropriate molecules and imaging modalities to assay gene expression. We now report the design, synthesis, and evaluation of novel enhanced reporter molecules, which reveal lacZ gene expression: trifluoromethylated aryl beta-D-galactopyranosides. A series of five molecular structures were screened in solution and with stably transfected lacZ expressing human MCF7 breast cancer cells in vitro. p-Trifluoromethyl-o-nitrophenyl beta-D-galactopyranoside (PCF(3)ONPG) was found to exhibit valuable properties including a single (19)F NMR signal, stability in aqueous solution and with wild type cells, but a chemical shift response to enzyme cleavage (Deltadelta=1.14 ppm) in breast cancer cells transfected to stably express lacZ.
Yu
Jianxin
J
Department of Radiology, The University of Texas Southwestern Medical Center at Dallas, Dallas, TX, USA.
Liu
Li
L
Kodibagkar
Vikram D
VD
Cui
Weina
W
Mason
Ralph P
RP
D005461:Fluorine; D005690:Galactose / Q000737:chemistry*; D017930:Genes, Reporter; D009682:Magnetic Resonance Spectroscopy; D001616:beta-Galactosidase / Q000235:genetics / Q000378:metabolism*
D005461:Fluorine; D001616:beta-Galactosidase; D005690:Galactose