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Peroxidase proximity selection to identify aptamers targeting a subcellular location.
Brandon Wilbanks1, William Beimers1,2, Maria Dugan1,3
1Department of Biochemistry and Molecular Biology, Mayo Clinic, Rochester, MN 55905, USA.
PNAS Nexus
|May 30, 2023
Summary
Researchers developed a new method, peroxidase proximity selection (PPS), to create DNA aptamers for targeted delivery into specific cellular compartments. This approach enables the delivery of functional cargos like antibodies into endosomes.
Area of Science:
- Nanomedicine
- Molecular Biology
- Biotechnology
Background:
- Efficient delivery of functional cargos into specific subcellular compartments remains a challenge in nanomedicine.
- Systematic Evolution of Ligands by EXponential enrichment (SELEX) identifies DNA aptamers for target recognition, but subcellular targeting is difficult.
Purpose of the Study:
- To develop a generalizable subcellular SELEX approach for aptamer selection.
- To identify DNA aptamers capable of targeting specific subcellular locations within living cells.
Main Methods:
- Peroxidase proximity selection (PPS) was developed as a subcellular SELEX method.
- Engineered ascorbate peroxidase APEX2 was used for local biotinylation of DNA aptamers within cells.
- Aptamers were selected for their ability to access the cytoplasm and specific organelles.
Main Results:
- DNA aptamers were discovered that preferentially enter endosomes via macropinocytosis.
- A subset of these aptamers demonstrated access to the cytoplasm, interacting with APEX2.
- One selected aptamer successfully delivered an IgG antibody into endosomes.
Conclusions:
- Peroxidase proximity selection (PPS) is a viable strategy for isolating DNA aptamers with subcellular targeting capabilities.
- This method advances the potential for targeted intracellular delivery of therapeutic cargos.
- The identified aptamers show promise for applications in targeted drug and protein delivery.
