A qPCR assay for measuring the post-integrational DNA repair in HIV-1 replication

Andrey N. Anisenko (Coresponding Author), Ekaterina S. Knyazhanskaya, Maria G. Isaguliants, Marina B. Gottikh

Research output: Contribution to journalArticlepeer-review

5 Citations (Scopus)


The post-integrational gap repair is a critical and poorly studied stage of the lentiviral life cycle. It might be performed by various cellular DNA repair pathways but the exact mechanism of the repair process has not yet been described. One of the reasons for that is the lack of a functional quantitative assay that could precisely measure the amount of integrated viral DNA that has completed the post-integrational gap repair stage. Here, we present an approach that is based on a widely used Alu-specific PCR for the estimation of integrated viral DNA but includes several steps that allow discrimination between integrated-repaired and integrated-unrepaired viral DNA forms. We used the approach for the estimation of the kinetics of gap repair in a viral vector system and showed that the gap repair process starts at 17 h post infection and lasts 10 more hours. We also showed that the addition of Nu7441 – a small molecule inhibitor of DNA-breaks sensor kinase in the non-homologous end joining DNA repair pathway – specifically inhibits the gap repair process while having no influence on the integration itself.

Original languageEnglish
Pages (from-to)12-19
Number of pages8
JournalJournal of Virological Methods
Publication statusPublished - Dec 2018
Externally publishedYes


  • DNA repair
  • DNA-PK
  • DNA-PKcs
  • HIV
  • Integration
  • qPCR

Field of Science*

  • 3.1 Basic medicine
  • 1.6 Biological sciences

Publication Type*

  • 1.1. Scientific article indexed in Web of Science and/or Scopus database


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