G4-SLSELEX-Seq-driven discovery of a G4-specific targeting L-RNA aptamer with unique structural features.

Area of Science:

• Molecular Biology
• Biochemistry
• Genetics

Background:

• DNA G-quadruplexes (dG4s) are unique nucleic acid structures with significant biological roles.
• Aptamers are valuable tools for studying and targeting nucleic acid structures.
• Developing specific aptamers for dG4s remains a challenge.

Purpose of the Study:

• To develop a novel aptamer selection strategy for DNA G-quadruplexes (dG4s).
• To identify and characterize L-RNA aptamers with high affinity and specificity for dG4s.
• To investigate the potential of aptamers in disrupting dG4-protein interactions.

Main Methods:

• Integration of sequence-guided library design with G4-SELEX-Seq.
• Selection and characterization of L-RNA aptamers against various dG4 structures.
• Affinity measurements using techniques like Surface Plasmon Resonance (SPR).
• Assessment of aptamer's ability to disrupt dG4-protein binding.

Main Results:

• The integrated approach successfully yielded aptamers targeting dG4s.
• Identified L-Apt.G3, the first L-RNA aptamer with low nanomolar binding affinity for dG4s.
• L-Apt.G3 demonstrated dual-specificity for both parallel and antiparallel dG4 conformations.
• L-Apt.G3 effectively competed with proteins binding to dG4s.

Conclusions:

• Sequence-guided library design enhances aptamer selection for complex targets like dG4s.
• L-Apt.G3 represents a novel and potent dG4-targeting aptamer with therapeutic and diagnostic potential.
• This study provides a powerful platform for developing aptamers against G-quadruplex structures.