DNA-Mediated Cellular Delivery of Functional Enzymes

Area of Science:

• Biomaterials Science
• Enzyme Engineering
• Nanotechnology

Background:

• Protein delivery into cells is crucial for therapeutic and research applications.
• Existing methods often suffer from low efficiency and require high protein concentrations.
• Functional proteins need to retain their native structure and activity after modification.

Purpose of the Study:

• To develop a new class of protein transfection materials.
• To enhance cellular uptake and intracellular activity of enzymes.
• To investigate the impact of oligonucleotide functionalization on protein properties.

Main Methods:

• Chemically modifying a functional protein core (β-galactosidase) with a dense shell of oligonucleotides (DNA strands).
• Characterizing the structural integrity and catalytic function of the modified protein.
• Assessing cellular uptake efficiency and intracellular enzyme activity compared to unmodified protein.

Main Results:

• Successfully created DNA-functionalized β-galactosidase that retains native structure and catalytic ability.
• Achieved up to a ~280-fold enhancement in cellular uptake.
• Enabled effective intracellular catalysis at working concentrations as low as 100 pM.

Conclusions:

• Oligonucleotide-functionalized proteins represent a promising new class of transfection materials.
• This strategy significantly improves protein delivery and intracellular function.
• The approach allows for highly efficient enzyme delivery at significantly reduced concentrations.