Note

Novel NMR approach to assessing gene transfection: 4-fluoro-2-nitrophenyl-β-D-galactopyranoside as a prototype reporter molecule for β-galactosidase

  1. Weina Cui1,†,
  2. Pieter Otten1,†,
  3. Yingming Li2,
  4. Kenneth S. Koeneman2,
  5. Jianxin Yu1,
  6. Ralph P. Mason1,*

Article first published online: 25 FEB 2004

DOI: 10.1002/mrm.10719

Issue

Magnetic Resonance in Medicine

Magnetic Resonance in Medicine

Volume 51, Issue 3, pages 616–620, March 2004

Additional Information

How to Cite

Cui, W., Otten, P., Li, Y., Koeneman, K. S., Yu, J. and Mason, R. P. (2004), Novel NMR approach to assessing gene transfection: 4-fluoro-2-nitrophenyl-β-D-galactopyranoside as a prototype reporter molecule for β-galactosidase. Magn. Reson. Med., 51: 616–620. doi: 10.1002/mrm.10719

Author Information

  1. 1

    Department of Radiology, University of Texas Southwestern Medical Center at Dallas, Dallas, Texas

  2. 2

    Department of Urology, University of Texas Southwestern Medical Center at Dallas, Dallas, Texas

  1. †

    Weina Cui and Pieter Otten contributed equally to this work

*University of Texas Southwestern Medical Center at Dallas, 5323 Harry Hines Blvd., Dallas, TX 75390-9058

Publication History

  1. Issue published online: 25 FEB 2004
  2. Article first published online: 25 FEB 2004
  3. Manuscript Revised: 9 OCT 2003
  4. Manuscript Accepted: 9 OCT 2003
  5. Manuscript Received: 17 APR 2003

Funded by

  • Department of Defense. Grant Numbers: BC980020 DAMD17-99-1-9381, BC022001 DAMD17-03-1-0343-01, DAMD17-01-1-0107
  • NIH Cancer Imaging Program. Grant Number: P20 CA 86354

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Keywords:

  • β-galactosidase;
  • gene reporter molecule;
  • lacZ;
  • 19F NMR;
  • nitrophenyl-β-D-galactopyranoside;
  • prostate cancer

Abstract

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 19F 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 19F 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.

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