Multivalent DNA recognition by self-assembled clusters: deciphering structural effects by fragments screening and...

Area of Science:

• Biochemistry
• Molecular Biology
• Gene Therapy

Background:

• Effective gene delivery vehicles are crucial for therapeutic oligonucleotide applications.
• Self-assembly processes offer a modular approach to creating complex biomolecular structures.

Purpose of the Study:

• To screen molecular fragments for creating low-molecular-weight clusters capable of binding therapeutic oligonucleotides.
• To evaluate the potential of these clusters for small interfering RNA (siRNA) delivery and gene silencing.

Main Methods:

• Utilized a one-pot fragment screening methodology based on chemoselective ligation.
• Assessed DNA binding using fluorescence displacement and gel electrophoresis assays.
• Evaluated siRNA transfection efficiency in MCF7 human breast cancer cells.

Main Results:

• Identified structural parameters influencing DNA binding affinity.
• Discovered specific biomolecular clusters with effective oligonucleotide complexation capabilities.
• Demonstrated successful delivery of a 21-mer siRNA into MCF7 cells via a developed cluster.

Conclusions:

• The developed modular methodology enables efficient screening for DNA-binding clusters.
• One identified biomolecular cluster shows promise for targeted siRNA delivery and gene therapy applications.
• Effective gene knockdown was achieved in cancer cells, highlighting the potential of this gene delivery system.