Efficient cellular uptake of click nucleic acid modified proteins.

Area of Science:

• Biotechnology
• Cell Biology
• Drug Delivery

Background:

• Efficient intracellular delivery of biomacromolecules like proteins is crucial for therapeutic applications but remains a significant challenge.
• Current methods often struggle with low efficiency and potential cytotoxicity, limiting their clinical utility.

Purpose of the Study:

• To investigate the efficacy of conjugating proteins to Click Nucleic Acids (CNAs) for enhanced intracellular delivery.
• To elucidate the cellular mechanisms involved in the uptake of CNA-conjugated proteins.

Main Methods:

• Conjugation of mScarlet (a red fluorescent protein) to Click Nucleic Acid (CNA).
• Assessment of cellular uptake in MDA-MB-468 cells compared to native proteins and DNA-conjugated proteins.
• Investigation of the role of metabolic activity, scavenger receptors, and caveolae-mediated endocytosis.
• Co-localization studies using anti-scavenger receptor antibodies.

Main Results:

• CNA-conjugated mScarlet exhibited significantly higher cell uptake than native proteins or DNA-conjugated proteins.
• The uptake process was confirmed to be metabolically dependent.
• Evidence suggests that scavenger receptors and caveolae-mediated endocytosis are key pathways for cellular entry.

Conclusions:

• Conjugation with Click Nucleic Acids (CNAs) represents a promising strategy for improving protein intracellular delivery.
• Scavenger receptors play a significant role in the uptake mechanism of CNA-modified proteins.
• This approach holds potential for advancing therapeutic protein delivery systems.