In Vitro Selection of Diversely Functionalized Aptamers

Area of Science:

• Biochemistry
• Molecular Biology
• Biotechnology

Background:

• Nucleic acid aptamers are crucial in molecular recognition and diagnostics.
• Developing aptamers with enhanced affinity and specificity remains a key challenge.
• Functional group modifications can significantly impact aptamer performance.

Purpose of the Study:

• To evolve a diversely functionalized nucleic acid aptamer for human alpha-thrombin.
• To investigate the role of functional group diversity in aptamer evolution.
• To establish a novel method for creating highly functionalized aptamers.

Main Methods:

• Utilized T4 DNA ligase-catalyzed DNA templated oligonucleotide polymerization.
• Employed a 256-membered ANNNN comonomer library with 16 functionalized sublibraries.
• Applied in vitro selection techniques for aptamer discovery.

Main Results:

• Successfully raised a highly functionalized aptamer with a dissociation constant of 1.6 nM for human alpha-thrombin.
• Demonstrated that functional modifications were essential for achieving high binding affinity.
• Confirmed the aptamer's selectivity for thrombin over other targets.

Conclusions:

• This study presents the most differentially functionalized nucleic acid aptamer discovered via in vitro selection.
• The developed method enables exploration of functional group dependence in nucleic acid polymer activity.
• The findings pave the way for designing next-generation aptamers with tailored properties.