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 Kodibagkar, Jian Xin Yu, Ralph P. Mason

Research output: Contribution to journalArticle

49 Citations (Scopus)

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 2007
Externally publishedYes

Fingerprint

Galactosidases
Lac Operon
prostatic neoplasms
enzymatic hydrolysis
Galactose
Heterografts
Gene expression
galactosidases
Tumors
Prostate
Hydrolysis
Prostatic Neoplasms
Nuclear magnetic resonance
Gene Expression
Gene therapy
gene expression
Molecular biology
Gene encoding
mice
Transgenes

Keywords

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

ASJC Scopus subject areas

  • Agricultural and Biological Sciences (miscellaneous)
  • Biochemistry, Genetics and Molecular Biology(all)
  • Biochemistry
  • Cell Biology
  • Medicine(all)

Cite this

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

In: FASEB Journal, Vol. 21, No. 9, 07.2007, p. 2014-2019.

Research output: Contribution to journalArticle

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