title: Stabilization and photochemical regulation of antisense agents through PEGylation.
abstract: Oligonucleotides are effective tools for the regulation of gene expression in cell culture and model organisms, most importantly through antisense mechanisms. Due to the inherent instability of DNA antisense agents, various modifications have been introduced to increase the efficacy of oligonucleotides, including phosphorothioate DNA, locked nucleic acids, peptide nucleic acids, and others. Here, we present antisense agent stabilization through conjugation of a poly(ethylene glycol) (PEG) group to a DNA oligonucleotide. By employing a photocleavable linker between the PEG group and the antisense agent, we were able to achieve light-induced deactivation of antisense activity. The bioconjugated PEG group provides stability to the DNA antisense agent without affecting its native function of silencing gene expression via RNase H-catalyzed mRNA degradation. Once irradiated with UV light of 365 nm, the PEG group is cleaved from the antisense agent leaving the DNA unprotected and open for degradation by endogenous nucleases, thereby restoring gene expression. By using a photocleavable PEG group (PhotoPEG), antisense activity can be regulated with high spatial and temporal resolution, paving the way for precise regulation of gene expression in biological systems.
authors.affiliation: Department of Chemistry, North Carolina State University, Raleigh, North Carolina 27695, United States.
authors.forename: Jeane M
authors.initials: JM
authors.lastname: Govan
authors.forename: Andrew L
authors.initials: AL
authors.lastname: McIver
authors.forename: Alexander
authors.initials: A
authors.lastname: Deiters
mesh_terms: D000818:Animals; D016373:DNA, Antisense / Q000138:chemical synthesis / Q000737:chemistry* / Q000235:genetics* / Q000378:metabolism; D005786:Gene Expression Regulation; D057809:HEK293 Cells; D006801:Humans; D008164:Luminescent Proteins / Q000235:genetics; D051379:Mice; D041681:NIH 3T3 Cells; D009691:Nucleic Acid Denaturation; D010782:Photolysis; D011092:Polyethylene Glycols / Q000737:chemistry*; D016914:Ribonuclease H / Q000378:metabolism; D014466:Ultraviolet Rays
chemical_list: D016373:DNA, Antisense; D008164:Luminescent Proteins; C413663:fluorescent protein 583; D011092:Polyethylene Glycols; D016914:Ribonuclease H