Off‐Target Effects Related to the Phosphorothioate Modification of Nucleic Acids

Citations Over TimeTop 17% of 2010 papers

Abstract

Phosphorothioate antisense oligonucleotides have been widely used in clinical studies for rational sequence-specific gene silencing. However, several sequence-unspecific off-target effects have been recently described for this compound class. In contrast to siRNA-mediated knockdown of the same gene, the bcl-2-targeted oblimersen (Genasense, G3139) downregulates a number of proteins involved in apoptotic resistance and several glycolytic enzymes in 607B human melanoma cells. Regardless of their target, phosphorothioate-modified antisense and siRNA compounds, but not oligonucleotides with a phosphodiester backbone, resulted in a similar impact on the proteome. Unspecifically downregulated proteins include cancer markers involved in apoptotic resistance and endoplasmatic reticulum (ER) stress such as the 78 kDa glucose regulated protein (GRP 78), protein disulfide isomerase A3 (PDIA3, GRP 58), calumenin, and galectin-1, as well as the glycolytic enzymes triose phosphate isomerase, glyceraldehyde phosphodehydrogenase, and phosphoglycerate mutase. The depletion of the glycolytic enzymes is reflected by a decrease in L-lactate production, indicating a partial reversal of the Warburg effect. Compared with other phosphorothioate oligonucleotides, oblimersen generally led to a more pronounced effect both in terms of the number of influenced proteins and the extent of downregulation, suggesting a synergistic effect of Bcl-2 downregulation.

Related Papers

• → Triosephosphate isomerase is a common crystallization contaminant of soluble His-tagged proteins produced inEscherichia coli(2013)8 cited
• → Triosephosphate isomerase from young and old Turbatrix aceti(1976)39 cited
• → Phosphorus-31 nuclear magnetic resonance spectroscopy reveals two conformational forms of chloroacetol phosphate-bound triosephosphate isomerase(1990)7 cited
• → Phenotypes of Phosphoglucose isomerase, phosphoglucose mutase and general protein in some freshwater molluscs from Basrah, Iraq(1985)
• Isozymes and In Vivo Activity of Triosephosphate Isomerase(1974)