Peroxidase proximity selection to identify aptamers targeting a subcellular location.

Autor:	Wilbanks B; Department of Biochemistry and Molecular Biology, Mayo Clinic, Rochester, MN 55905, USA., Beimers W; Department of Biochemistry and Molecular Biology, Mayo Clinic, Rochester, MN 55905, USA.; Present address: Department of Biochemistry, University of Wisconsin-Madison, Madison, WI 53706, USA., Dugan M; Department of Biochemistry and Molecular Biology, Mayo Clinic, Rochester, MN 55905, USA.; Present address: Department of Chemistry, Iowa State University, Ames, IA 50011, USA., Weiskittel T; Department of Molecular Pharmacology and Experimental Therapeutics, Mayo Clinic, Rochester, MN 55901, USA., Maher LJ 3rd; Department of Biochemistry and Molecular Biology, Mayo Clinic, Rochester, MN 55905, USA.
Jazyk:	angličtina
Zdroj:	PNAS nexus [PNAS Nexus] 2023 May 04; Vol. 2 (5), pp. pgad151. Date of Electronic Publication: 2023 May 04 (Print Publication: 2023).
DOI:	10.1093/pnasnexus/pgad151
Abstrakt:	The efficient and specific delivery of functional cargos such as small-molecule drugs, proteins, or nucleic acids across lipid membranes and into subcellular compartments is a significant unmet need in nanomedicine and molecular biology. Systematic Evolution of Ligands by EXponential enrichment (SELEX) exploits vast combinatorial nucleic acid libraries to identify short, nonimmunogenic single-stranded DNA molecules (aptamers) capable of recognizing specific targets based on their 3D structures and molecular interactions. While SELEX has previously been applied to identify aptamers that bind specific cell types or gain cellular uptake, selection of aptamers capable of carrying cargos to specific subcellular compartments is challenging. Here, we describe peroxidase proximity selection (PPS), a generalizable subcellular SELEX approach. We implement local expression of engineered ascorbate peroxidase APEX2 to biotinylate naked DNA aptamers capable of gaining access to the cytoplasm of living cells without assistance. We discovered DNA aptamers that are preferentially taken up into endosomes by macropinocytosis, with a fraction apparently accessing APEX2 in the cytoplasm. One of these selected aptamers is capable of endosomal delivery of an IgG antibody. (© The Author(s) 2023. Published by Oxford University Press on behalf of National Academy of Sciences.)
Databáze:	MEDLINE
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