Stabilization and Photochemical Regulation of Antisense Agents through PEGylation
Stabilization and Photochemical Regulation of Antisense Agents through PEGylation
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DOI：
10.1021/bc200411n
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发表时间：
2011-10-01
影响因子：
4.7
通讯作者：
Deiters, Alexander
中科院分区：
文献类型：
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作者：
Govan, Jeane M.;McIver, Andrew L.;Deiters, Alexander
寡核苷酸是调节细胞培养和模型生物中基因表达的有效工具，最重要的是通过反义机制。由于DNA反义剂固有的不稳定性，人们引入了各种修饰来提高寡核苷酸的功效，包括硫代磷酸酯DNA、锁定核酸、肽核酸等。在这里，我们通过将聚乙二醇 (PEG) 基团与 DNA 寡核苷酸缀合来实现反义剂的稳定性。通过在 PEG 基团和反义剂之间使用可光裂解的连接体，我们能够实现光诱导的反义活性失活。生物共轭 PEG 基团为 DNA 反义剂提供稳定性，而不影响其通过 RNase H 催化 mRNA 降解沉默基因表达的天然功能。一旦用 365 nm 的紫外线照射，PEG 基团就会从反义剂上裂解下来，使 DNA 不受保护并开放供内源核酸酶降解，从而恢复基因表达。通过使用可光裂解的 PEG 基团 (PhotoPEG)，可以以高空间和时间分辨率调节反义活性，为生物系统中基因表达的精确调节铺平道路。
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.