Posted on the website on 3 May 2005.
Control of DNA Hybridization with Photocleavable Adducts¶
Article first published online: 30 APR 2007
Photochemistry and Photobiology
Volume 81, Issue 4, pages 953–959, July 2005
How to Cite
Ghosn, B., Haselton, F. R., Gee, K. R. and Monroe, W. T. (2005), Control of DNA Hybridization with Photocleavable Adducts. Photochemistry and Photobiology, 81: 953–959. doi: 10.1111/j.1751-1097.2005.tb01469.x
- Issue published online: 30 APR 2007
- Article first published online: 30 APR 2007
- Received 15 November 2004; accepted 27 April 2005
Previous reports have shown that 1-(4,5-dimethoxy-2-nitro-phenyl)ethyl ester (DMNPE) adducts coupled to DNA plasmids block transcription in vitro and in vivo until removed with light. In this report, we explore the use of DMNPE to control DNA hybridization. We found that DMNPE-caged oligonucleotides have changed spectrophotometric and elec-trophoretic properties that can be restored with light exposure. Caged oligonucleotides have slower electrophoretic mobility than noncaged oligonucleotides and caged oligonucleotides exposed to light. Effects of caging on hybridization were assessed in a fluorescence-based assay using a 20mer caged DNA oligonucleotide complementary to a 30mer molecular beacon. Fluorescence results indicate that hybridization is reduced and subsequently restored by light. Subsequent gel shift assays confirmed these results. Hybridization activity of caged oligonucleotides with an average of 14–16 DMNPE adducts per oligonucleotide was 14% of noncaged control oligonucleotides and after 365 nm photolysis, increased to nearly 80% of controls. Spectrophotometric characterization of caged oligonucleotides exposed to light and then filtered to remove the released DMNPE adducts indicates two to four attached cage groups remaining following photoactivation. These results suggest that this light-based technology can be used as a tool for the spatial and temporal regulation of hybridization-based DNA bioactivity.