Inducible regulating homologous recombination enables precise genome editing in Pichia pastoris without perturbing cellular fitness

• Published in: Trends in biotechnology (Regular ed.) Vol. 43; no. 6; pp. 1385 - 1402
• Main Authors: Bai, Fan, Cai, Peng, Yao, Lun, Shen, Yiwei, Li, Yunxia, Zhou, Yongjin J.
• Format: Journal Article
• Language: English
• Published: England Elsevier Ltd 01.06.2025; Elsevier Limited
• Subjects: adverse effects; Biosynthesis; biotechnology; Cell cycle; Cell growth; CRISPR; CRISPR-Cas9; dynamic regulation; Editing; Efficiency; fatty alcohols; Fitness; Fungal Proteins - genetics; Fungal Proteins - metabolism; Gene deletion; Gene Editing - methods; Gene Expression Regulation, Fungal; Genes; Genetic engineering; genome assembly; Genome editing; Genomes; Glucose; Homologous recombination; Homologous Recombination - genetics; Industrial applications; Internal Medicine; Kinases; Komagataella pastoris; methanol; Natural products; Pichia - genetics; Pichia pastoris; Proteins; Rad52 DNA Repair and Recombination Protein - genetics; Rad52 protein; RNA polymerase; Saccharomycetales - genetics; tetracycline; Yeast; yeasts; dynamic regulation; genome editing; Komagataella pastoris; Pichia pastoris; homologous recombination; CRISPR-Cas9

A tetracycline repressor protein (TetR)/tetO2 inducible system was constructed and optimized in Pichia pastoris.The TetR/tetO2 inducible system dynamically enhanced homologous recombination (HR) for precise editing of multiple genes.Dynamically enhancing HR did not compromise cellular growth or prod...