Evolution of Functionally Enhanced α-l-Threofuranosyl Nucleic Acid Aptamers

ACS Synth Biol. 2021 Nov 19;10(11):3190-3199. doi: 10.1021/acssynbio.1c00481. Epub 2021 Nov 5.

Abstract

Synthetic genetic polymers (xeno-nucleic acids, XNAs) have the potential to transition aptamers from laboratory tools to therapeutic agents, but additional functionality is needed to compete with antibodies. Here, we describe the evolution of a biologically stable artificial genetic system composed of α-l-threofuranosyl nucleic acid (TNA) that facilitates the production of backbone- and base-modified aptamers termed "threomers" that function as high quality protein capture reagents. Threomers were discovered against two prototypical protein targets implicated in human diseases through a combination of in vitro selection and next-generation sequencing using uracil nucleotides that are uniformly equipped with aromatic side chains commonly found in the paratope of antibody-antigen crystal structures. Kinetic measurements reveal that the side chain modifications are critical for generating threomers with slow off-rate binding kinetics. These findings expand the chemical space of evolvable non-natural genetic systems to include functional groups that enhance protein target binding by mimicking the structural properties of traditional antibodies.

Keywords: aptamers; in vitro selection; machine learning; therapeutics; threose nucleic acid; xeno-nucleic acids.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Antibodies / chemistry
  • Aptamers, Nucleotide / chemistry*
  • Kinetics
  • Nucleic Acids / chemistry*
  • Polymers / chemistry*
  • Proteins / chemistry
  • Tetroses / chemistry*

Substances

  • Antibodies
  • Aptamers, Nucleotide
  • Nucleic Acids
  • Polymers
  • Proteins
  • Tetroses