Kimoto, M., Matsunaga, K. and Hirao, I. ( 2017). Evolving aptamers with unnatural base pairs. Current Protocols in Chemical Biology, 9, 315– 339. doi: 10.1002/cpch.31
A novel technology, genetic alphabet expansion, has rapidly advanced through the successful creation of unnatural base pairs that function as a third base pair in replication. Recently, genetic alphabet expansion has been applied to some practical areas. Among them, the application to DNA aptamer generation is a good example of the broad utility of this technology. A hydrophobic unnatural base pair, Ds–Px, which exhibits high fidelity in replication as a third base pair, has been applied to an evolutionary engineering method called SELEX (Systematic Evolution of Ligands by EXponential enrichment) to generate DNA aptamers that bind to targets. A few Ds bases in DNA aptamers significantly increase the binding affinity to targets, enabling the use of DNA aptamers as an alternative to antibodies. This protocol describes the ExSELEX (genetic alphabet Expansion for SELEX) method to generate Ds-containing DNA aptamers.
This work was supported by the Institute of Bioengineering and Nanotechnology (Biomedical Research Council, Agency for Science, Technology and Research, Singapore).
This is the peer reviewed version of the following article: Kimoto, M., Matsunaga, K. and Hirao, I. ( 2017). Evolving aptamers with unnatural base pairs. Current Protocols in Chemical Biology, 9, 315– 339. doi: 10.1002/cpch.31, which has been published in final form at https://doi.org/10.1002/cpch.31. This article may be used for non-commercial purposes in accordance with Wiley Terms and Conditions for Use of Self-Archived Versions.