Isolation and characterization of high affinity aptamers against DNA polymerase iota

Andrei V. Lakhin, Andrei A. Kazakov, Alena V. Makarova, Youri I Pavlov, Anna S. Efremova, Stanislav I. Shram, Viacheslav Z. Tarantul, Leonid V. Gening

Research output: Contribution to journalArticle

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Abstract

Human DNA-polymerase iota (Pol ι) is an extremely error-prone enzyme and the fidelity depends on the sequence context of the template. Using the in vitro systematic evolution of ligands by exponential enrichment (SELEX) procedure, we obtained an oligoribonucleotide with a high affinity to human Pol ι, named aptamer IKL5. We determined its dissociation constant with homogenous preparation of Pol ι and predicted its putative secondary structure. The aptamer IKL5 specifically inhibits DNA-polymerase activity of the purified enzyme Pol ι, but did not inhibit the DNA-polymerase activities of human DNA polymerases beta and kappa. IKL5 suppressed the error-prone DNA-polymerase activity of Pol ι also in cellular extracts of the tumor cell line SKOV-3. The aptamer IKL5 is useful for studies of the biological role of Pol ι and as a potential drug to suppress the increase of the activity of this enzyme in malignant cells.

Original languageEnglish (US)
Pages (from-to)49-57
Number of pages9
JournalNucleic Acid Therapeutics
Volume22
Issue number1
DOIs
StatePublished - Feb 1 2012

Fingerprint

DNA-Directed DNA Polymerase
SELEX Aptamer Technique
Enzymes
Oligoribonucleotides
DNA Polymerase beta
Tumor Cell Line
Tumors
Cells
Ligands
Pharmaceutical Preparations
DNA polymerase iota

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Medicine
  • Molecular Biology
  • Genetics
  • Drug Discovery

Cite this

Lakhin, A. V., Kazakov, A. A., Makarova, A. V., Pavlov, Y. I., Efremova, A. S., Shram, S. I., ... Gening, L. V. (2012). Isolation and characterization of high affinity aptamers against DNA polymerase iota. Nucleic Acid Therapeutics, 22(1), 49-57. https://doi.org/10.1089/nat.2011.0324

Isolation and characterization of high affinity aptamers against DNA polymerase iota. / Lakhin, Andrei V.; Kazakov, Andrei A.; Makarova, Alena V.; Pavlov, Youri I; Efremova, Anna S.; Shram, Stanislav I.; Tarantul, Viacheslav Z.; Gening, Leonid V.

In: Nucleic Acid Therapeutics, Vol. 22, No. 1, 01.02.2012, p. 49-57.

Research output: Contribution to journalArticle

Lakhin, AV, Kazakov, AA, Makarova, AV, Pavlov, YI, Efremova, AS, Shram, SI, Tarantul, VZ & Gening, LV 2012, 'Isolation and characterization of high affinity aptamers against DNA polymerase iota', Nucleic Acid Therapeutics, vol. 22, no. 1, pp. 49-57. https://doi.org/10.1089/nat.2011.0324
Lakhin, Andrei V. ; Kazakov, Andrei A. ; Makarova, Alena V. ; Pavlov, Youri I ; Efremova, Anna S. ; Shram, Stanislav I. ; Tarantul, Viacheslav Z. ; Gening, Leonid V. / Isolation and characterization of high affinity aptamers against DNA polymerase iota. In: Nucleic Acid Therapeutics. 2012 ; Vol. 22, No. 1. pp. 49-57.
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