Evolution of Cross-Linking Aptamers for Long-Lasting Membrane Protein Labeling

Area of Science:

• Biochemistry
• Molecular Biology
• Chemical Biology

Background:

• Membrane protein research is crucial and growing.
• Aptamers offer structural flexibility and efficient screening for ligand discovery.
• Investigating membrane proteins requires robust tools.

Purpose of the Study:

• To develop a novel in vitro evolution strategy for covalent aptamers.
• To leverage fluorine-sulfur exchange (SuFEx) chemistry for aptamer development.
• To create aptamers for specific membrane protein targeting and cell classification.

Main Methods:

• In vitro evolution strategy.
• Fluorine-sulfur exchange (SuFEx) chemistry.
• Screening for aptamers with high specificity to membrane proteins.

Main Results:

• Developed covalent aptamers, termed cross-linking aptamers (xl-aptamers).
• Achieved aptamers with sustained and specific binding affinity for target proteins.
• Demonstrated efficient cell classification using the developed aptamers.

Conclusions:

• The novel SuFEx-based strategy is effective for developing specific aptamers.
• Covalent aptamers show significant potential for single-cell and spatial proteomics.
• This method advances membrane protein research and cell analysis.