Hervey John R D, Freund Niklas, Houlihan Gillian, Dhaliwal Gurpreet, Holliger Philipp, Taylor Alexander I
Cambridge Institute of Therapeutic Immunology & Infectious Disease (CITIID), University of Cambridge Cambridge CB2 0AW UK
Medical Research Council Laboratory of Molecular Biology Cambridge CB2 0QH UK
RSC Chem Biol. 2022 Aug 30;3(10):1209-1215. doi: 10.1039/d2cb00035k. eCollection 2022 Oct 5.
Functional nucleic acids can be evolved using cycles of selection and amplification, starting from diverse-sequence libraries, which are typically restricted to natural or partially-modified polymer chemistries. Here, we describe the efficient DNA-templated synthesis and reverse transcription of libraries entirely composed of serum nuclease resistant alternative nucleic acid chemistries validated in nucleic acid therapeutics; locked nucleic acid (LNA), 2'--methyl-RNA (2'OMe-RNA), or mixtures of the two. We evaluate yield and diversity of synthesised libraries and measure the aggregate error rate of a selection cycle. We find that in addition to pure 2'--methyl-RNA and LNA, several 2'OMe-RNA/LNA blends seem suitable and promising for discovery of biostable functional nucleic acids for biomedical applications.
功能核酸可以从多样序列文库开始，通过选择和扩增循环进行进化，这些文库通常限于天然或部分修饰的聚合物化学。在这里，我们描述了完全由在核酸治疗中得到验证的血清核酸酶抗性替代核酸化学组成的文库的高效DNA模板合成和逆转录；锁核酸（LNA）、2'-O-甲基-RNA（2'OMe-RNA）或两者的混合物。我们评估了合成文库的产量和多样性，并测量了一个选择循环的总错误率。我们发现，除了纯的2'-O-甲基-RNA和LNA外，几种2'OMe-RNA/LNA混合物似乎适用于并有望用于发现用于生物医学应用的生物稳定功能核酸。
