Fluorescence Labeling of GC-Rich DNA Matrix with Different Nucleotide Derivatives for Hybridization Analysis on a Biological Microarray

Capa

Citar

Texto integral

Acesso aberto Acesso aberto
Acesso é fechado Acesso está concedido
Acesso é fechado Somente assinantes

Resumo

We investigated the efficiency of labeling the GC-rich promoter region of the TERT gene in the human genome with derivatives of 5'-triphosphates of 2'-deoxyuridine (dU) and 2'-deoxycytidine (dC) containing cyanine dyes Cy5 and Cy7 as a fluorescent label. The effect of modified nucleotides on the efficiency of the polymerase chain reaction was evaluated by real-time PCR, and the extent of nucleotide incorporation into the PCR product was also determined. The efficiency of DNA matrix labeling was determined by the intensity of fluorescent signal during allele-specific hybridization on a biological microarray. The highest level of biochip cell fluorescence was observed for dU-Cy7 derivatives compared to dU- and dC-Cy5 derivatives. At the same time, in the case of GC-rich DNA matrix, the use of dC-Cy5 derivatives gave a fundamentally better result compared to dU-Cy5 derivative. Thus, modified Cy7 analogs capable of incorporation into DNA during PCR are less dependent on the GC composition of the DNA matrix and are more universal fluorescent labels for diagnostic purposes. Further application of modified Cy7 analogs in the development of laboratory-on-a-chip test systems seems to be the most promising.

Sobre autores

I. Barinova

Engelhardt Institute of Molecular Biology, Russian Academy of Sciences

Moscow, 119991 Russia

V. Shershov

Engelhardt Institute of Molecular Biology, Russian Academy of Sciences

Moscow, 119991 Russia

V. Varachev

Engelhardt Institute of Molecular Biology, Russian Academy of Sciences

Moscow, 119991 Russia

S. Surzhikov

Engelhardt Institute of Molecular Biology, Russian Academy of Sciences

Moscow, 119991 Russia

I. Grechishnikova

Engelhardt Institute of Molecular Biology, Russian Academy of Sciences

Moscow, 119991 Russia

O. Zasedateleva

Engelhardt Institute of Molecular Biology, Russian Academy of Sciences

Moscow, 119991 Russia

T. Nasedkina

Engelhardt Institute of Molecular Biology, Russian Academy of Sciences

Email: tanased06@rambler.ru
Moscow, 119991 Russia

A. Chudinov

Engelhardt Institute of Molecular Biology, Russian Academy of Sciences

Moscow, 119991 Russia

Bibliografia

  1. Stumpf A., Brandstetter T., Hübner J., Rühe J. // PLoS One. 2019. V. 14. P. e0225525. https://doi.org/10.1371/journal.pone.0225525
  2. Dullaert-de Boer M., Akkerman O.W., Vermeer M., Hess D.L.J., Kerstjens H.A.M., Anthony R.M., van der Werf T.S., van Soolingen D., van der Zanden A.G.M. // PLoS One. 2018. V. 13. P. e0190847. https://doi.org/10.1371/journal.pone.0190847
  3. Ikonnikova A.Y., Filippova M.A., Surzhikov S.A., Pozhitnova V.O., Kazakov R.E., Lisitsa T.S., Belkov S.A., Nasedkina T.V. // Drug Metab. Pers. Ther. 2020. V. 36. P. 33-40. https://doi.org/10.1515/dmpt-2020-0155
  4. Leevy W.M., Gammon S.T., Johnson J.R., Lampkins A.J., Jiang H., Marquez M., Piwnica-Worms D., Suckow M.A., Smith B.D. // Bioconj. Chem. 2008. V. 19. P. 686-692. https://doi.org/10.1021/bc700376v
  5. Шершов В.Е., Иконникова А.Ю., Василисков В.А., Лапа С.А., Мифтахов Р.А., Кузнецова В.Е., Чудинов А.В., Наседкина Т.В. // Биофизика. 2020. Т. 65. С. 865-871. https://doi.org/10.31857/S0006302920050038
  6. Фесенко Д.О., Гусейнов Т.О., Лапа С.А., Кузнецова В.Е., Шершов В.Е., Спицын М.А., Наседкина Т.В., Заседателев А.С., Чудинов А.В. // Мол. биология. 2018. Т. 52. С. 533-542. https://doi.org/10.7868/S0026898418030175
  7. Иконникова А.Ю., Шершов В.Е., Мороз Ю.В., Василисков В.А., Лапа С.А., Мифтахов Р.А., Кузнецова В.Е., Чудинов А.В., Наседкина Т.В. // Биофизика. 2021. Т. 66. С. 31-39. https://doi.org/10.31857/S000630292101004X
  8. Bell R.J., Rube H.T., Xavier-Magalhães A., Costa B.M., Mancini A., Song J.S., Costello J.F. // Mol. Cancer Res. 2016. V. 14. P. 315-323. https://doi.org/10.1158/1541-7786.MCR-16-0003
  9. Фесенко Д.О., Абрамов И.С., Шершов В.Е., Кузнецова В.Е., Суржиков С.А., Гречишникова И.В., Барский В.Е., Чудинов А.В., Наседкина Т.В. // Мол. биология. 2018. Т. 52. С. 997-1005. https://doi.org/10.1134/S0026898418060071
  10. Lapa S.A., Volkova O.S., Spitsyn M.A., Shershov V.E., Kuznetsova V.E., Guseinov T.O., Zasedatelev A.S., Chudinov A.V. // Russ. J. Bioorg. Chem. 2019. V. 45. P. 263-272. https://doi.org/10.1134/S1068162019040046
  11. Иконникова А.Ю., Лисица Т.С., Шершов В.Е., Спицын М.А., Гусейнов Т.О., Фесенко Д.О., Лапа С.А., Кузнецова В.Е., Заседателев А.С., Чудинов А.В., Наседкина Т.В. // Биофизика. 2017. Т. 62. С. 1093- 1098.
  12. Boyer A.E., Lipowska M., Zen J.-M., Patonay G. // Anal. Lett. 1992. V. 25. P. 415-428. https://doi.org/10.1080/00032719208016105
  13. Varachev V.O., Guskov D.A., Shekhtman A.P., Rogozhin D.V., Polyakov S.A., Zacedatelev A.S., Chudinov A.V., Nasedkina T.V. // Russ. J. Bioorg. Chem. 2023. V. 49. P. 1137-1142. https://doi.org/10.1134/S1068162023050205
  14. Varachev V., Shekhtman A., Guskov D., Rogozhin D., Zasedatelev A., Nasedkina T. // Diagnostics (Basel). 2024. V. 14. P. 200. https://doi.org/10.3390/diagnostics14020200

Arquivos suplementares

Arquivos suplementares
Ação
1. JATS XML
Baixar

Declaração de direitos autorais © Russian Academy of Sciences, 2025

Согласие на обработку персональных данных

 

Используя сайт https://journals.rcsi.science, я (далее – «Пользователь» или «Субъект персональных данных») даю согласие на обработку персональных данных на этом сайте (текст Согласия) и на обработку персональных данных с помощью сервиса «Яндекс.Метрика» (текст Согласия).