Single-step selection of bivalent aptamers validated by comparison with SELEX using high-throughput sequencing.

Area of Science:

• Biochemistry
• Molecular Biology
• Bioinformatics

Background:

• Nucleic acid aptamer selection typically involves multiple rounds of amplification and enzymatic transcription.
• Streamlining aptamer identification can significantly advance molecular diagnostics and therapeutics.

Purpose of the Study:

• To compare the efficacy of traditional SELEX (Systematic Evolution of Ligands by Exponential Enrichment) with single-step selection for identifying bivalent aptamers targeting thrombin.
• To evaluate the utility of next-generation sequencing and motif-finding informatics in simplifying aptamer discovery.

Main Methods:

• SELEX and single-step selection methods were employed to identify aptamers against thrombin.
• Next-generation sequencing and bioinformatics tools were used to analyze the selected aptamer pools.
• Motif-finding algorithms were applied to identify conserved sequences and potential aptamers.

Main Results:

• Both SELEX and single-step selection identified similar aptamers targeting thrombin.
• Next-generation sequencing and motif-finding informatics proved effective in analyzing aptamer pools from both selection methods.
• The findings suggest that advanced informatics can reduce the number of selection rounds required.

Conclusions:

• Next-generation sequencing and motif-finding informatics offer a powerful approach to simplify nucleic acid aptamer selection.
• These methods have the potential to reduce the time and resources needed for aptamer discovery.
• This advancement could accelerate the development of aptamer-based technologies.