Yeast Base Editing Service

The Yeast Base Editing Service offers an advanced, highly precise tool for making targeted Single-Nucleotide Polymorphism (SNP) changes in the yeast genome (Saccharomyces cerevisiae, Pichia pastoris, etc.). Base editing utilizes a modified CRISPR-Cas9 system—a catalytically impaired Cas9 fused to a base-modifying enzyme (a deaminase)—which chemically converts one DNA base into another (e.g., C>T or A>G) without creating a Double-Strand Break (DSB).

CD Biosynsis’s base editing platform significantly improves the efficiency and precision of point mutation introduction compared to traditional CRISPR-Cas9 combined with Homologous Recombination (HR). This service is essential for protein engineering (site-directed mutagenesis), fine-tuning metabolic pathways (altering enzyme kinetics), and introducing stop codons for highly efficient gene knockout. We deliver fully verified yeast strains with the exact desired base pair change, accelerating strain optimization and functional genomics studies.

Highlights

Key advantages of utilizing Base Editing for yeast genome modification:

• Ultra-High Precision: Enables specific C>T/G>A and A>G/T>C base conversions at the target site with minimal unintended edits.
• No Double-Strand Breaks: Bypasses the need for DNA repair mechanisms (like HR or NHEJ), which often lead to unintended insertions or deletions (indels).
• Increased Efficiency: Significantly higher efficiency for introducing point mutations compared to traditional HR-based methods, especially in non-model yeast.
• Scarless Modification: Introduces the point mutation directly into the genome without the need for large donor DNA templates or selection markers.

Editing Applications

Applications benefiting from single-base precision in yeast:

Protein Directed Evolution

Introducing specific missense mutations (altering amino acids) in target enzymes to enhance activity, stability, or specificity.

Metabolic Fine-Tuning

Altering promoter or ribosome binding sites via point mutation to subtly adjust gene expression levels and balance metabolic flux.

High-Efficiency Gene Knockout

Introducing a premature stop codon (via C>T or A>G conversion) at the start of a gene to ensure complete, stable knockout.

Functional Genomics

Creating precise SNP variants found in natural yeast populations or pathological strains for functional characterization.

Key Features & Precision

Technical features of our optimized Yeast Base Editing platform:

Cytosine Base Editors (CBE)

Enables the conversion of C:G base pairs to T:A base pairs (C>T conversion) with high specificity.

Adenine Base Editors (ABE)

Enables the conversion of A:T base pairs to G:C base pairs (A>G conversion), expanding the editing scope.

Optimized Editing Window

gRNA design is optimized to position the target base within the narrow editing window for maximum conversion efficiency.

Minimized Indel Formation

The absence of DSBs ensures extremely low frequency of insertions or deletions (indels), maintaining genomic integrity.

Full Sequence Validation

Rigorous verification of the entire target region via Sanger sequencing to confirm the desired base change and check for off-target edits.

Workflow

Our systematic workflow for precise yeast base editing:

• Target and Editor Selection: Select the target gene and the appropriate base editor (CBE or ABE) based on the desired base change.
• gRNA Design and Cloning: Design high-specificity gRNAs to position the target base within the editor's activity window.
• Yeast Transformation: Deliver the base editor and gRNA expression plasmids into the target yeast strain.
• Editing and Selection: Allow time for the base editing activity, followed by selection for successfully edited clones.
• Verification and Curing: Sequence the target locus to confirm the precise base change. Cure the final strain of the editor and gRNA plasmids to ensure stability.
• Delivery: Delivery of the verified, stable, base-edited yeast strain and complete sequence data.

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

• Guaranteed SNP: Commitment to deliver a verified yeast clone with the exact single-base change at the specified locus.
• High Editing Purity: Guaranteed purity of the edited clones, with extremely low rates of indel formation or mixed edits.
• Stable Final Strain: Assurance that the final strain is cured of editing components, maintaining the stability of the chromosomal edit.
• Comprehensive Verification: Rigorous sequencing validation to confirm the target edit and check for potential unintended modifications in the target region.