Smart Nucleic Acid Chaperones: Phase-Separating Intrinsically Disordered Proteins for Accelerating DNA Hybridization...

Area of Science:

• Molecular Biology
• Biotechnology
• Nanotechnology

Background:

• Nucleic acid (NA) hybridization is fundamental to molecular biology and NA nanotechnologies.
• NA chaperones catalyze NA structure formation via base-pairing.
• Intrinsically disordered proteins (IDPs) exhibit dynamic behavior and phase separation.

Purpose of the Study:

• Develop smart NA chaperones (SNACs) by merging NA chaperoning function with IDP-driven phase separation.
• Investigate SNACs' ability to control NA hybridization and reaction kinetics.
• Explore SNACs as tools for NA nanotechnologies and nanoassembly.

Main Methods:

• Engineered fusion proteins combining NA chaperones and IDPs.
• Designed IDPs with inherent NA chaperoning capabilities.
• Assessed SNACs' impact on nucleic acid strand annealing (SA) and toehold-mediated strand displacement (TMSD) kinetics.

Main Results:

• SNACs significantly enhanced the kinetics of both SA and TMSD reactions.
• Engineered elastin-like protein-NA-binding domain fusions formed protein-DNA coacervates that accelerated SA.
• SNACs accelerated TMSD in both soluble and phase-separated states.

Conclusions:

• SNACs are effective tools for controlling NA hybridization reactions.
• Leveraging phase separation with SNACs enhances reaction kinetics.
• SNACs show potential as versatile nanoassemblers for complex NA systems.