Wang Xiaolong, Xian Jingnü, Chen Gang, Peng Haibo
Department of Biotechnology, College of Marine Life Sciences, Ocean University of China, Qingdao 266003, Shandong, China.
Sheng Wu Gong Cheng Xue Bao. 2018 May 25;34(5):664-675. doi: 10.13345/j.cjb.170401.
Oligonucleotides are widely used as effective tools to regulate gene expression and drugs for targeted gene therapy. Therefore, they are potentially useful for the treatment of viral, tumor and hereditary diseases. Therapeutic oligonucleotides include antisense oligonucleotide, small interference RNA (siRNA), Ribozyme, DNAzyme, anti-gene, CpG, decoy and aptamer. Therapeutic oligonucleotides usually carry certain modifications, such as phosphorothioates, fluoro or locked nucleic acids, to enhance the stability and specificity, and reduce the side-effects, because natural oligonucleotides have poor stability in vivo, low specificity and side effects. Now oligonucleotides are usually manufactured by chemical synthesis, with low purity and high cost. Here, we review a novel thermocyclic reaction for the amplification of oligonucleotides, referred to as Polymerase-endonuclease Amplification Reaction (PEAR) catalyzed by two thermostable enzymes. PEAR is simple, efficient, and stable. Comparing with traditional chemical synthesis, PEAR-based enzymatic production of oligonucleotides could be a robust alternative method for the large-scale production of therapeutic or non-therapeutic oligonucleotides.
寡核苷酸作为调节基因表达的有效工具和靶向基因治疗的药物被广泛应用。因此，它们在治疗病毒、肿瘤和遗传性疾病方面具有潜在用途。治疗性寡核苷酸包括反义寡核苷酸、小干扰RNA（siRNA）、核酶、脱氧核酶、反基因、CpG、诱饵和适体。治疗性寡核苷酸通常带有某些修饰，如硫代磷酸酯、氟或锁核酸，以提高稳定性和特异性，并减少副作用，因为天然寡核苷酸在体内稳定性差、特异性低且有副作用。目前寡核苷酸通常通过化学合成制备，纯度低且成本高。在此，我们综述了一种用于寡核苷酸扩增的新型热循环反应，称为由两种耐热酶催化的聚合酶-核酸内切酶扩增反应（PEAR）。PEAR简单、高效且稳定。与传统化学合成相比，基于PEAR的寡核苷酸酶促生产可能是大规模生产治疗性或非治疗性寡核苷酸的一种强大替代方法。
