CRISPR-Cas9 Genome Engineering

Yeast CRISPR-Cas9 Genome Editing Service

Our Yeast CRISPR-Cas9 Genome Editing Service delivers precise, sequence-confirmed modifications in Saccharomyces cerevisiae for research and engineering applications.

Custom gRNA design and validation
Cas9 and donor vector construction
Sequence-verified edited yeast clones

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Why Choose Our Yeast Editing Service?

We provide a streamlined, end-to-end CRISPR-Cas9 workflow for Saccharomyces cerevisiae, from guide design to validated clonal isolates. Our service supports gene knockouts, point mutations, and targeted insertions to accelerate your research and metabolic engineering projects.

Experienced in project-qualified systems as scoped and ploidies
Rigorous PCR and Sanger sequencing validation
Customizable donor design for precise edits

Gene Knockout

Point Mutation

Targeted Insertion

Our Approach

Systematic CRISPR-Cas9 Workflow for Yeast

We combine computational gRNA design, robust Cas9 expression, and efficient homologous recombination to deliver high-fidelity genome edits in Saccharomyces cerevisiae.

Custom gRNA Design & Validation

We analyze your target region using proprietary algorithms to select gRNAs with high on-target activity and minimal off-target potential. gRNAs are cloned into expression vectors and validated before transformation.

Off-target prediction via in silico tools
Cloning into Cas9 expression vector
Sequence verification of gRNA construct

Donor Vector Construction & HR

For precise edits requiring homology-directed repair, we design and construct donor vectors with homology arms tailored to your modification. For knockouts, indels are generated via non-homologous end joining.

Custom homology arm design (typically 300–800 bp)
Incorporation of desired mutation or tag
QC by restriction digest and sequencing

Streamlined Transformation

High-efficiency lithium acetate/PEG transformation into your selected Saccharomyces cerevisiae strain, with selection on appropriate auxotrophic or antibiotic markers.

Clonal Isolation & Screening

Individual clones are picked and screened by colony PCR using locus-specific primers. Positive clones are re-streaked to support clonal purity.

Sequence Verification

Sanger sequencing across the edited locus confirms the intended modification and absence of undesired mutations. Results are documented in a final report.

What You Receive

Project Deliverables

Each project includes verified material and comprehensive documentation to support your downstream applications.

Materials

≥1 validated yeast clone (glycerol stock or colony)
GRNA expression plasmid (if requested)
Cas9 expression plasmid (if supplied by us)
Donor vector construct (for HDR projects)

Documentation

Final project report with gRNA sequences and design rationale
Sanger sequencing chromatograms and alignments
PCR screening gel images
QC summary including homogeneity assessment

Service Scope

Customizable Parameters

Projects are tailored to your specific requirements. Below are typical parameters that can be adjusted per consultation.

Parameter	Typical project scope
Target organism	Saccharomyces cerevisiae (common lab strains; other strains evaluated upon request)
Edit type	Scoped per project after consultation
Number of gRNAs	1–4 gRNAs per target as needed for efficiency; pool or multiplex options
Donor design	Custom homology arms (length based on edit); synthetic ssODN or plasmid donor
Selection marker	Auxotrophic markers (e.g., URA3, LEU2) or antibiotic resistance (e.g., G418, hygromycin)
Ploidy consideration	Haploid or diploid; screening protocol adjusted for multiple alleles
Clone validation	Scoped per project after consultation
Off-target analysis	In silico prediction; optional targeted amplicon sequencing of top off-target sites

How It Works

Project Workflow

A straightforward five-step process from consultation to delivery of validated clones.

Project Design & gRNA Selection

Share your target gene and desired edit. Our team designs gRNAs using computational tools, evaluates off-target potential, and selects the best candidates. You approve the design before we proceed.

Vector Construction

We clone the gRNA(s) into a Cas9 expression vector and, if required, build a donor repair template with appropriate homology arms. All constructs are sequence-verified before transformation.

Yeast Transformation & Selection

Competent yeast cells (your strain or a standard lab strain) are transformed with the Cas9/gRNA plasmid and donor (if applicable).

Clonal Screening & Picking

Individual colonies are picked and screened by colony PCR using primers flanking the target locus. Positive clones are re-streaked to support monoclonality and re-screened.

Validation & Delivery

Confirmed clones are expanded, genomic DNA is extracted, and Sanger sequencing is performed across the edited region. A final report with sequencing results and a stock of validated clones is delivered to you.

Applications

Where Yeast Genome Editing Adds Value

Our service supports a wide range of research and industrial goals in Saccharomyces cerevisiae.

Functional Genomics

Knockout or tag endogenous genes to study function, regulation, and protein localization in a model eukaryote.

Gene–phenotype association studies
Epitope tagging for protein tracking
Promoter replacement for expression analysis

Pathway Optimization

Disrupt competing pathways, introduce heterologous enzymes, or fine-tune flux to evaluate production of metabolites, biofuels, or chemicals.

Knockout of byproduct pathways
Integration of biosynthetic gene clusters
Point mutations to alter enzyme activity

Genome-Scale Rewiring

Perform multiplex edits to reshape yeast metabolism or create synthetic circuits for novel functions.

Multiple gene knockouts in a single project
Precise integration of synthetic cassettes
Replacement of regulatory elements

Frequently Asked Questions

Common Questions About Yeast CRISPR Editing

Answers to help you plan your project.

Projects from design to delivery of validated clones typically take a timeline confirmed during project scoping, depending on the complexity of the edit and number of targets. Timeline is confirmed in the project scope.
Yes. Our screening protocol can be adjusted for diploid or aneuploid strains to identify biallelic or multi-copy edits. Please discuss your strain during project intake.
We use colony PCR followed by Sanger sequencing across the target locus. Sequencing data is compared to the wild-type reference to confirm the intended modification and check for off-target mutations in the amplified region.
Yes, sequential or multiplex editing is possible. We can design projects for two or more loci with appropriate selection marker recycling or use of multiple gRNAs. Complexity and timeline scale accordingly.
You receive validated yeast clones (glycerol stock or liquid culture), a detailed project report including gRNA sequences, PCR gel images, Sanger sequencing chromatograms, and alignments. Plasmids can be provided upon request.

Scientific Literature

References

Waldrip ZJ, Jenjaroenpun P, DeYoung O, et al. Genome-wide Cas9 binding specificity in Saccharomyces cerevisiae. PeerJ. 2020;8:e9442. View on PubMed
Laughery MF, Hunter T, Brown A, et al. New vectors for simple and streamlined CRISPR-Cas9 genome editing in Saccharomyces cerevisiae. Yeast (Chichester, England). 2015;32(12):711-20. View on PubMed
Molinet J, Urbina K, Villegas C, et al. A Saccharomyces eubayanus haploid resource for research studies. Scientific reports. 2022;12(1):5976. View on PubMed
DiCarlo JE, Norville JE, Mali P, et al. Genome engineering in Saccharomyces cerevisiae using CRISPR-Cas systems. Nucleic acids research. 2013;41(7):4336-43. View on PubMed
Bindal G, Amlinger L, Lundgren M, et al. Type I-E CRISPR-Cas System as a Defense System in Saccharomyces cerevisiae. mSphere. 2022;7(3):e0003822. View on PubMed

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