Pichia pastoris genome editing

Service Introduction

Pichia pastoris is a non-pathogenic, obligate aerobic haploid budding yeast and one of the most outstanding "protein production factories" in the field of molecular biology. It is a methylotrophic yeast capable of utilizing methanol as its sole carbon source and energy source. Punter Biotech employs third-generation editing technologies, represented by the CRISPR/Cas9 system, which offer high efficiency and simplicity for targeted modifications of the Pichia pastoris genome, including gene knockout, foreign sequence insertion, and point mutation introduction, thereby enabling functional optimization of strains and reconstruction of metabolic pathways.

Service Advantages

▶Extensive Experience: Possesses extensive expertise in Pichia pastoris gene editing, with a seasoned team capable of efficiently constructing Pichia pastoris mutants.

▶Quality Assurance: All gene editing sites are rigorously sequenced and validated to ensure product quality.

Service Process

Article Case

Combinatorial optimization of CRISPR/Cas9 expression enables precision genome engineering in the methylotrophic yeast Pichia pastoris

academic journal：Accepted Manuscript

(B) Cells were transformed with plasmids (based on design #82, Tab. 1) bearing HsCAS9, GUT1-gRNA2 and a gRNA to target AOX1 (gRNA1, gRNA2 or gRNA3) and stamped onto BMD1, BMM1 and glycerol (BMG1) containing agar plates Depending on the gRNA combination used multiplexing efficiencies up to 69% were obtained. Mean values and standard deviations of biological triplicates are shown.(C) Representative experimental data of the phenotypic characterization of the experiments in (B) are shown (CRISPR/Cas9 plasmid bearing the gRNAs AOX1-gRNA2 and GUT1-gRNA2). As controls the P. pastoris wildtype strain (WT), mutS (ΔAOX1) and a ΔGUT1 strain were used .

Applicable Fields

▶Metabolic Engineering and Compound Synthesis: Capable of efficiently synthesizing various high-value compounds, and even performing complex multi-gene editing to reconstruct metabolic pathways.

▶ Protein Expression and Pharmaceutical Industry: More effectively addresses the inherent limitations of the host organism, enhancing both yield and product quality.

▶Strain Functional Optimization: In addition to direct product production, gene editing is also frequently employed to systematically enhance the intrinsic properties of Pichia pastoris chassis cells.

▶ Fundamental Scientific Research: By editing specific genes, researchers can investigate their functions and elucidate life mechanisms such as growth, metabolism, and stress responses in Pichia pastoris.