DNA aptamer generation by ExSELEX using genetic alphabet expansion with a mini-hairpin DNA stabilization method

Ichiro Hirao; Michiko Kimoto; Kyung Hyun Lee

doi:10.1016/j.biochi.2017.09.007

DNA aptamer generation by ExSELEX using genetic alphabet expansion with a mini-hairpin DNA stabilization method

Biochimie. 2018 Feb:145:15-21. doi: 10.1016/j.biochi.2017.09.007. Epub 2017 Sep 13.

Authors

Ichiro Hirao¹, Michiko Kimoto², Kyung Hyun Lee²

Affiliations

¹ Institute of Bioengineering and Nanotechnology, 31 Biopolis Way, The Nanos, #09-01, Singapore, 138669, Singapore. Electronic address: ichiro@ibn.a-star.edu.sg.
² Institute of Bioengineering and Nanotechnology, 31 Biopolis Way, The Nanos, #09-01, Singapore, 138669, Singapore.

PMID: 28916151
DOI: 10.1016/j.biochi.2017.09.007

Abstract

A novel aptamer generation method to greatly augment the affinity and stability of DNA aptamers was developed by genetic alphabet expansion combined with mini-hairpin DNA technology. The genetic alphabet expansion increases the physicochemical and structural diversities of DNA aptamers by introducing extra components, unnatural bases, as a fifth base, allowing for the enhancement of DNA aptamer affinities. Furthermore, the mini-hairpin DNA technology stabilizes DNA aptamers against nuclease digestion and thermal denaturation, by introducing an extraordinarily stable mini-hairpin DNA containing a GCGAAGC sequence. This novel method provides stabilized high-affinity DNA aptamers for diagnostic and therapeutic applications.

Keywords: Aptamer; ExSELEX; Genetic alphabet expansion; Mini-hairpin DNA; Unnatural base pair.

Publication types

Review

MeSH terms

Aptamers, Nucleotide / chemical synthesis*
Aptamers, Nucleotide / chemistry*
Aptamers, Nucleotide / genetics*
Aptamers, Nucleotide / therapeutic use
Humans
Nucleic Acid Conformation*
SELEX Aptamer Technique / methods*

Substances

Aptamers, Nucleotide