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Characterizing Oligonucleotide Uptake in Cultured Cells: A Case Study Using AS1411 Aptamer

  • Elsa M. Reyes-Reyes
  • Paula J. BatesEmail author
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 2036)

Abstract

Oligonucleotides can be designed or evolved to bind to specific DNA, RNA, protein, or small molecule targets and thereby alter the biological function of the target. The therapeutic potential of oligonucleotides targeted to intracellular molecules will depend largely on their ability to be taken up by the cells of interest, as well as their subsequent subcellular distribution. Here we describe methods to characterize the extent and mechanism of cellular uptake of AS1411, an aptamer oligonucleotide that has progressed to human clinical trials and which is also widely used by researchers as a cancer-targeting ligand.

Key words

Oligonucleotide AS1411 Aptamer G-quadruplex Nucleolin Endocytosis Macropinocytosis Cancer Flow cytometry Confocal microscopy 

Notes

Acknowledgments

The work described was funded by NIH grant R01CA122383 (Bates). The authors are named coinventors on issued patents or patent applications pertaining to AS1411

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Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2019

Authors and Affiliations

  1. 1.Division of Pulmonary, Allergy, Critical Care, and Sleep MedicineUniversity of Arizona College of MedicineTucsonUSA
  2. 2.Department of Cellular and Molecular MedicineUniversity of Arizona Cancer CenterTucsonUSA
  3. 3.Department of MedicineUniversity of LouisvilleLouisvilleUSA
  4. 4.Department of BiochemistryUniversity of LouisvilleLouisvilleUSA
  5. 5.Experimental Therapeutics Program of the James Graham Brown Cancer CenterUniversity of LouisvilleLouisvilleUSA

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