19F-NMR detection of lacZ gene expression via the enzymic hydrolysis of 2-fluoro-4-nitrophenyl β-D-galactopyranoside in vivo in PC3 prostate tumor xenografts in the mouse

Li Liu, Vikram D. Kodibagkar, Jian Xin Yu, Ralph P. Mason

Research output: Contribution to journalArticle

49 Scopus citations

Abstract

Gene therapy shows promise for treating prostate cancer and has been evaluated in several clinical trials. A major challenge that remains is to establish a method for verifying transgene activity in situ. The lacZ gene encoding β-galactosidase historically has been the most popular reporter gene for molecular biology. We have designed a 19F NMR approach to reveal lacZ gene expression by assessing β-galactosidase (β-gal) activity in vivo. The substrate 2-fluoro-4-nitrophenyl β-D- galactopyranoside (OFPNPG) is readily hydrolyzed by β-gal with a corresponding decrease in the 19F-NMR signal from OFPNPG and the appearance of a new signal shifted 4-6 ppm upfield from the aglycone 2-fluoro-4-nitrophenol (OFPNP). We report proof of principle in cultures of PC3 prostate cancer cells using 19F NMR spectroscopy and 19F chemical shift imaging. More importantly, we demonstrate for the first time the ability to differentiate wild-type and lacZ-expressing prostate tumor xenografts in mice using this approach.

Original languageEnglish (US)
Pages (from-to)2014-2019
Number of pages6
JournalFASEB Journal
Volume21
Issue number9
DOIs
StatePublished - Jul 1 2007

Keywords

  • F CSI
  • F MRS
  • Gene reporter
  • β-galactosidase

ASJC Scopus subject areas

  • Biotechnology
  • Biochemistry
  • Molecular Biology
  • Genetics

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