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July 13, 2026Journal of Biological EngineeringOpen Access

An advanced genome engineering platform for Komagataella phaffii enabling versatile in vivo DNA assembly and multiplex integration

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Authors

YCYujung ChoiKyung Hee UniversityAKA Leum KimKyung Hee UniversityNKNam Kyu KangSeoul National University

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Implication

Randomized trial reveals improved DNA assembly and multipathway integration in Komagataella phaffii, suggesting a versatile platform for synthetic biology applications.

Key Points

  • The goal is to develop a robust genome engineering platform for Komagataella phaffii to enhance DNA assembly and integration capabilities.
  • Created dKR chassis by deleting Ku70 and overexpressing RAD52 to enhance homologous recombination.
  • Utilized CRISPR/Cas9-mediated editing for high-efficiency multigene integration.
  • Implemented tRNA-ribozyme system for simultaneous integration at multiple genomic sites.
  • Increased integration efficiency from 15.3% to 68.1% using short 50-bp homology arms.
  • Achieved 86.5% integration efficiency for nine linear DNA fragments with vector-free assembly.
  • Produced β-carotene in a functional biosynthetic pathway while maintaining normal growth and productivity.

Cite This Study

Choi et al. (2026) studied this question.

synapsesocial.com/papers/6a548105475c38bf615a5825https://doi.org/10.1186/s13036-026-00731-z
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Also Consider

Synapse has enriched 5 closely related papers on similar clinical questions. Consider them for comparative context:

  1. 1Efficient CRISPR‐mediated C‐to‐T base editing in Komagataella phaffii2024 · 10 citations
  2. 2CRISPR‐Cas9 mediated genome editing of Kluyveromyces marxianus for iterative, multiplexed gene disruption and pathway integration2024 · 11 citations
  3. 3Comparison of CRISPR-MAD7 and CRISPR-Cas9 for Gene Disruptions in Komagataella phaffii2024 · 8 citations
  4. 4Efficient and markerless gene integration with SlugCas9-HF in Kluyveromyces marxianus2024 · 20 citations
  5. 5Metabolic engineering of Komagataella phaffii for enhanced production of heterologous proteins and small molecules: key strategies and future directions2026 · 1 citations