Using the Flp Recombinase to Induce Site-Specific Protein-DNA Nicks

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Updated: Feb 13, 2026

Principles of Site-Specific Recombinase SSR Technology
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Principles of Site-Specific Recombinase SSR Technology

Published on: May 29, 2008

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Kristoffer P Jakobsen1, Lotte Bjergbæk1

  • 1Aarhus University, Aarhus, Denmark.

Methods in Enzymology
|March 11, 2018
PubMed

Insights

Researchers created the Flp-nick system to study protein-linked DNA nicks (PDNs), a common DNA damage. This system enables detailed analysis of PDN repair pathways, crucial for maintaining genome stability.

Area of Science:

  • Molecular Biology
  • Genetics
  • Biochemistry

Background:

  • DNA Topoisomerase I (Top1) activity generates protein-linked DNA nicks (PDNs), a frequent DNA insult.
  • If unrepaired, PDNs can convert to double-strand breaks (DSBs) during replication, leading to genome instability.
  • Understanding PDN repair is critical for cell fate and genome integrity.

Purpose of the Study:

  • To develop a novel cellular system for generating and studying single, site-specific PDNs.
  • To investigate the molecular mechanisms underlying PDN repair pathways.
  • To provide a versatile tool for analyzing DNA repair in various genomic contexts.

Main Methods:

  • Development of the Flp-nick system in Saccharomyces cerevisiae.
  • Utilizing a catalytically active but non-ligating Flp recombinase mutant.
  • Inducing a single, site-specific PDN at a defined genomic locus.
  • Employing molecular analysis to study repair pathway dynamics.

Main Results:

  • The Flp-nick system successfully generates site-specific PDNs mimicking Top1-induced damage.
  • The system allows for detailed molecular investigation of PDN repair processes.
  • Demonstrated the utility of the system for studying repair in specific genomic environments.

Conclusions:

  • The Flp-nick system is a powerful tool for dissecting PDN repair mechanisms.
  • This system facilitates the study of genome stability maintenance.
  • Offers a flexible platform for future DNA repair research.
Keywords:
Break-induced replicationDouble-strand break repairNick repairProtein–DNA adductReplication damageTopoisomerase I

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