Imaging beta-galactosidase activity using 19F chemical shift imaging of LacZ gene-reporter molecule 2-fluoro-4-nitrophenol-beta-D-galactopyranoside.
2-Fluoro-4-nitrophenol-beta-D-galactopyranoside (OFPNPG) belongs to a novel class of NMR active molecules (fluoroaryl-beta-D-galactopyranosides), which are highly responsive to the action of beta-galactosidase (beta-gal). OFPNPG has a single 19F peak (-55 ppm relative to aqueous sodium trifluoroacetate). Upon cleavage by beta-gal, the pH sensitive aglycone 2-fluoro-4-nitrophenol (OFPNP) is observed at a chemical shift of -59 to -61 ppm. The chemical shift response is sufficient to observe beta-gal activity using chemical shift imaging (CSI). 19F CSI studies of enzyme activity and lacZ gene expression in 9L-glioma and MCF7 breast cancer cells are presented, providing further evidence for the utility of OFPNPG as a gene-reporter molecule for future in vivo studies.
Kodibagkar
Vikram D
VD
Department of Radiology, University of Texas Southwestern Medical Center at Dallas, Dallas, TX 75390-9058, USA.
Yu
Jianxin
J
Liu
Li
L
Hetherington
Hoby P
HP
Mason
Ralph P
RP
D001943:Breast Neoplasms / Q000235:genetics / Q000378:metabolism; D004926:Escherichia coli / Q000201:enzymology; D005462:Fluorine Radioisotopes; D017930:Genes, Reporter / Q000235:genetics*; D005910:Glioma / Q000235:genetics / Q000378:metabolism; D006801:Humans; D007763:Lac Operon / Q000235:genetics*; D009682:Magnetic Resonance Spectroscopy / Q000379:methods*; D009598:Nitrophenylgalactosides; D011401:Promoter Regions, Genetic / Q000235:genetics; D019275:Radiopharmaceuticals; D014162:Transfection; D001616:beta-Galactosidase / Q000235:genetics / Q000378:metabolism*
C521953:2-fluoro-4-nitrophenyl galactopyranoside; D005462:Fluorine Radioisotopes; D019275:Radiopharmaceuticals; D009598:Nitrophenylgalactosides; D001616:beta-Galactosidase