Premium
Novel NMR approach to assessing gene transfection: 4‐fluoro‐2‐nitrophenyl‐β‐ D ‐galactopyranoside as a prototype reporter molecule for β‐galactosidase
Author(s) -
Cui Weina,
Otten Pieter,
Li Yingming,
Koeneman Kenneth S.,
Yu Jianxin,
Mason Ralph P.
Publication year - 2004
Publication title -
magnetic resonance in medicine
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.696
H-Index - 225
eISSN - 1522-2594
pISSN - 0740-3194
DOI - 10.1002/mrm.10719
Subject(s) - transfection , reporter gene , chemistry , gene expression , in vivo , gene , enzyme , fluorine 19 nmr , gene product , molecule , small molecule , biochemistry , nuclear magnetic resonance spectroscopy , microbiology and biotechnology , biophysics , stereochemistry , biology , genetics , organic chemistry
Gene therapy holds great promise for the treatment of diverse diseases. However, widespread implementation is hindered by difficulties in assessing the success of transfection in terms of spatial extent, gene expression, and longevity of expression. The development of noninvasive reporter techniques based on appropriate molecules and imaging modalities may help to assay gene expression. 4‐Fluoro‐2‐nitrophenyl‐β‐ D ‐galactopyranoside (PFONPG) is a novel prototype NMR‐sensitive molecule, which is highly responsive to the action of β‐galactosidase (β‐gal), the product of the lacZ gene. The molecule is stable in solution and with respect to wild‐type cells, but the enzyme causes very rapid liberation of the aglycone, accompanied by color formation and a 19 F NMR chemical shift of 5–10 ppm, depending on pH. Since the product is pH‐sensitive, this opens the possibility for direct pH determinations at the site of enzyme activity. Molecular and 19 F NMR characteristics of PFONPG in solution, blood, and prostate tumor cells are presented. This prototype molecule facilitates a novel approach for assaying gene activity in vivo. Magn Reson Med 51:616–620, 2004. © 2004 Wiley‐Liss, Inc.