Gene therapy has emerged as a promising strategy for treatment of various diseases, but there is a pressing need for the development of non-invasive reporter techniques based on appropriate molecules and imaging modalities to assay gene expression. We now report the design, synthesis, and evaluation of novel enhanced reporter molecules, which reveal lacZ gene expression: trifluoromethylated aryl β-d-galactopyranosides. A series of five molecular structures were screened in solution and with stably transfected lacZ expressing human MCF7 breast cancer cells in vitro. p-Trifluoromethyl-o- nitrophenyl β-d-galactopyranoside (PCF3ONPG) was found to exhibit valuable properties including a single 19F NMR signal, stability in aqueous solution and with wild type cells, but a chemical shift response to enzyme cleavage (Δδ = 1.14 ppm) in breast cancer cells transfected to stably express lacZ.
Bibliographical noteFunding Information:
This work was supported by grants from the DOD Breast Cancer Initiative IDEA award DAMD17-03-1-0343-01 and the Cancer Imaging Program, NIH P20 CA 86354 (pre-ICMIC). NMR experiments were conducted at the Mary Nell and Ralph B. Rogers NMR Center, an NIH BTRP facility #P41-RR02584.
- F NMR
- Reporter gene