ssDNA Modification on Lipid Carriers Utilizing ssDNA-Conjugated Fusion Protein with Cholesterol-Binding Ability

Area of Science:

• Biotechnology
• Nanotechnology
• Drug Delivery

Background:

• Lipid carriers (liposomes, exosomes) are key for drug and nucleic acid delivery.
• Surface functionalization with ligands improves targeted delivery.
• DNA aptamers offer high specificity and affinity for targeting applications.

Purpose of the Study:

• To develop an improved method for modifying lipid carriers with single-stranded DNA (ssDNA).
• To utilize a novel cholesterol-binding fusion protein (Rep-ALOD4) for ssDNA conjugation.
• To demonstrate cell-selective targeting using ssDNA-modified lipid carriers.

Main Methods:

• Engineered a fusion protein (Rep-ALOD4) combining ssDNA conjugation (Rep) and cholesterol-binding (ALOD4) domains.
• Applied Rep-ALOD4 to modify liposomes with >30 wt% cholesterol.
• Validated cell-selective targeting with ssDNA-aptamer-modified liposomes and exosomes.

Main Results:

• Successfully achieved ssDNA modification on cholesterol-rich liposomes using Rep-ALOD4.
• Demonstrated effective cell-selective targeting with the ssDNA-modified lipid carriers.
• Confirmed the functional applicability of ssDNA-displaying Rep-ALOD4 for targeting.

Conclusions:

• The Rep-ALOD4 platform provides an effective strategy for ssDNA modification of lipid carriers.
• This method significantly advances targeted drug delivery applications.
• The approach holds potential for future development in nanomedicine.