Yeast CRISPR-Cas9 Genome Editing Services

Highlights

Key advantages of utilizing CRISPR-Cas9 for Yeast Genome Editing:

• High HR Efficiency: Yeast’s strong homologous recombination pathway makes it naturally suited for precise, template-driven gene insertions and replacements.
• Multiplex Editing: Capability to target and modify multiple genes simultaneously in a single transformation round, accelerating complex pathway optimization.
• Scalable and Stable: Allows for chromosomal integration of entire synthetic pathways, ensuring superior genetic stability compared to unstable plasmids.
• Rapid Iteration: Significantly faster and more reliable than traditional methods (e.g., homologous recombination using selection markers) for pathway construction.

Editing Applications

Comprehensive genome editing solutions for yeast systems:

Gene Knockout and Deletion

Precise deletion of native genes to remove competing metabolic pathways or undesirable byproduct formation.

Gene Knock-In and Insertion

Stable, scarless chromosomal integration of large biosynthetic genes or entire gene clusters (pathway integration).

Promoter Engineering and Tuning

Precise replacement of native promoters with stronger, weaker, or inducible variants to control gene expression levels.

Allele Replacement and SNP Editing

Correction or introduction of specific single nucleotide polymorphisms (SNPs) for protein engineering or disease modeling.

Key Features & Efficiency

Technical features ensuring successful and precise yeast genome modifications:

Optimized gRNA Design

Proprietary algorithms for guide RNA design to minimize off-target effects and maximize on-target cleavage efficiency.

Seamless Donor Integration

Design of short or long homologous repair templates for efficient, high-fidelity integration without unwanted sequences.

High Multiplexing Capacity

Proven ability to edit up to 10 distinct genetic loci simultaneously in a single round for rapid metabolic pathway construction.

Versatile Host Compatibility

Expertise in editing various industrial yeast strains, including S. cerevisiae, P. pastoris, and Yarrowia lipolytica.

Full Sequence Verification

Verification of all edited loci via PCR amplification and Sanger sequencing to confirm successful, on-target modification.

Workflow

Our systematic workflow for high-precision yeast genome editing:

• Design and Template Synthesis: Design of gRNAs and synthesis of the DNA donor templates (for knock-ins or replacements).
• Plasmid Construction and Delivery: Assembly of CRISPR-Cas9/gRNA expression cassettes and preparation for yeast transformation.
• Yeast Transformation: Highly optimized transformation protocols to deliver the editing components efficiently into the target yeast host.
• Selection and Clone Isolation: Selection of successfully edited clones using transient markers or counter-selection strategies.
• Verification and Curing: Molecular verification (PCR and sequencing) of the edited loci, followed by removal of the Cas9 plasmid (curing).
• Delivery: Delivery of the verified, genetically stable, marker-free yeast strain and complete sequence data.

We provide essential assurance for high-quality yeast genome editing outcomes:

• Guaranteed Edit: Commitment to deliver a verified yeast clone with the specified genetic modification (KO, KI, or replacement).
• Sequence Fidelity: Guaranteed correct sequence of the inserted or modified region and confirmed on-target editing.
• Genetic Stability: Assurance that the final strain is cured of editing plasmids and is stable due to chromosomal integration.
• Multiplex Success: Experienced management of multiplex editing projects, ensuring simultaneous modification of complex pathways.