Khamissi Natalie, Korfmann Christopher, Chaudhry Areeba, Hili Ryan
Department of Chemistry, Centre for Research on Biomolecular Interactions, York University 4700 Keele Street Toronto ON M3J 1P3 Canada
Chem Sci. 2025 Mar 21. doi: 10.1039/d5sc00834d.
A method to enable the transliteration between various XNA-containing nucleic acids and canonical DNA is described. Using ligase-catalysed oligonucleotide polymerisation (LOOPER), we show that DNA can be used as a template to generate nucleic acids polymers comprising various levels of 2'-fluoro (2'-F), 2'-fluoro-arabinonucleic acid (FANA), 2'--methyl (2'-OMe), and Locked Nucleic Acids (LNA) in moderate yields. The fidelity and biases of the LOOPER process were studied in detail for the 2'-F system by developing a hairpin-based sequencing method, which showed fidelities exceeding 95% along with positional and sequence dependencies within the polymerised XNA-containing anticodons. Lastly, we show the ability of LOOPER to regenerate DNA from 2'-F, FANA, 2'-OMe, and LNA in moderate yield and in fidelities over 95%. Taken together, this study demonstrates the potential of LOOPER to serve as a platform for applications where the transliteration between XNA and DNA is needed, such as the evolution of XNA-containing nucleic acid polymers.
本文描述了一种实现各种含XNA的核酸与经典DNA之间相互转录的方法。利用连接酶催化的寡核苷酸聚合反应（LOOPER），我们证明DNA可用作模板，以中等产率生成包含不同水平的2'-氟（2'-F）、2'-氟阿拉伯核酸（FANA）、2'-O-甲基（2'-OMe）和锁核酸（LNA）的核酸聚合物。通过开发一种基于发夹的测序方法，我们详细研究了2'-F系统中LOOPER过程的保真度和偏向性，结果表明在聚合的含XNA反密码子内，保真度超过95%，同时存在位置和序列依赖性。最后，我们展示了LOOPER以中等产率和超过95%的保真度从2'-F、FANA、2'-OMe和LNA再生DNA的能力。综上所述，本研究证明了LOOPER作为一个平台的潜力，可用于需要XNA与DNA之间相互转录的应用，如含XNA核酸聚合物的进化。
