Seamless Gene Tagging by Endonuclease-Driven Homologous Recombination
2011

Seamless Gene Tagging in Yeast

publication Evidence: high

Author Information

Author(s): Anton Khmelinskii, Matthias Meurer, Nurlanbek Duishoev, Nicolas Delhomme, Michael Knop

Primary Institution: European Molecular Biology Laboratory (EMBL)

Hypothesis

Can a seamless gene tagging approach preserve endogenous gene regulation while allowing for high-throughput analysis?

Conclusion

The seamless gene tagging method preserves gene regulation and enables efficient high-throughput functional studies in yeast.

Supporting Evidence

  • The seamless tagging method allows for the preservation of upstream and downstream gene regulatory elements.
  • High-throughput strain construction is compatible with the seamless tagging approach.
  • Seamless tagging enables functional analysis of transcription antisense to open reading frames.

Takeaway

This study shows a new way to add tags to genes in yeast without messing up how those genes work, making it easier to study them.

Methodology

The study used an endonuclease-driven approach for seamless gene tagging in Saccharomyces cerevisiae, allowing for high-throughput strain construction and automated yeast genetics.

Limitations

The method may face challenges with ectopic homologous recombination in regions of high sequence similarity.

Digital Object Identifier (DOI)

10.1371/journal.pone.0023794

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