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CRISPR Genome Editing in Bacteria and Yeast

Metabolic engineering optimizes genetic and regulatory processes in microbes to increase the yield of key compounds, chemicals, and proteins for pharmaceutical and nutraceutical ingredient development. Microorganisms, such as E. coli and S. cerevisiae, are popular cell sources for these molecules, and with the recent CRISPR/Cas9 technology breakthrough, genome editing in bacteria and yeast has become more efficient and precise.

GenScript is pleased to introduce the Microbial genome editing service for knock-in, knock-out or gene replacement in bacteria (E. coli) and yeast (S. cerevisiae). Our service employs a two vector system to target up to three genes simultaneously, ensuring editing accuracy down to the base pair. In addition, to further improve efficacy in bacteria, our scientists have developed a novel λ Red-CRISPR/Cas9 system, which leverages the efficiency of traditional λ Red recombineering with the ease of CRISPR/Cas for seamless, targeted genome editing.

Advantages of GenScript’s CRISPR microbial gene editing system

CRISPR/Cas technology has quickly become one of the easiest and most effective genome editing systems, and when combined with λ Red recombineering has several advantages over the most common editing strategies:

• Seamless
• High efficiency: even in haploid and diploid yeast strains
• Multigene editing: can knock-out up to 3 genes simultaneously
• Precision down to the base pair
• Easy selection: no selectable marker is required

Service Details

*Standard gene synthesis services (SC1010) can be used for the KI gene insert. Click here to learn more about pricing and turnaround times.
**Pricing does not include the additional cost for gene synthesis, if required.

Case studies

Using CRISPR to generate a CAN1 knock-out S. cerevisiae strain Read more »

• CRISPR/Cas9 was used to delete a 400 bp sequence in the CAN1 gene to block gene expression (Figure 1).
• PCR of the recombinant strains yielded bands 400 bp shorter than the native strain, demonstrating successful gene knock-out (Figure 2). A 100% CAN1 knock-out success rate (10/10) was achieved using this system.

Using CRISPR to knock-in a 1kb fragment in the S. cerevisiae CAN1 gene Read more »

• The CRISPR system was used to insert a 1 kb fragment into the CAN1 gene to test KI efficiency (Figure 3).
• PCR of the recombinant strains yielded bands 1 kb longer than the native strain, indicating successful gene knock-in. A 100% success rate (5/5 strains) was achieved using CRISPR (Figure 4).

Related CRISPR Services

• Metabolic Pathway Assembly Service: optimize expression of multiple genes simultaneously.
• RBS Design: maximize translation of your target protein using our ribosome binding site design algorithm.
• CRISPR/Cas9 Technology Resources: Learn more about the CRISPR/Cas9 genome editing technology, find experimental protocols, or watch our free instructional webinars.

Quotations and Ordering

For quotation requests:

• Bacteria genome editing service: Call, email or submit an inquiry using our online request form.
• Yeast genome editing service: download and complete the yeast genome editing service request form.
  • Email the form or upload the form through our secure online request management server.

To order:

• Orders can be placed by phone, email, or fax with a formal PO (Purchase Order) or credit card.

Our customer service representatives are available 24 hours a day, Monday through Friday, to assist you.

Acknowledgements

We would like to acknowledge Professor Sheng Yang (Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences) for contributing the original plasmids for the microbial genome editing service. The plasmids were subsequently engineered by the GenScript team to meet the service needs.

Legal Statement of GenCRISPR Services and Products (Updated on July 28, 2015)*:

*The legal statement below applies to eukaryotic cells only (including yeast); currently, bacterial cells (E.coli) are exempt from these regulations.

1. GenCRISPR™ services and products are covered under US 8,697,359, US 8,771,945, US 8,795,965, US 8,865,406, US 8,871,445, US 8,889,356, US 8,889,418, US 8,895,308, US 8,906,616 and foreign equivalents and licensed from Broad Institute, Inc. Cambridge, Massachusetts.
2. The products and the reagents generated from these services shall be used as tools for research purposes, and shall exclude (a) any human or clinical use, including, without limitation, any administration into humans or any diagnostic or prognostic use, (b) any human germline modification, including modifying the DNA of human embryos or human reproductive cells, (c) any in vivo veterinary or livestock use, or (d) the manufacture, distribution, importation, exportation, transportation, sale, offer for sale, marketing, promotion or other exploitation or use of, or as, a testing service, therapeutic or diagnostic for humans or animals.
3. The purchase of the GenCRISPR Services and Products coveys to the purchaser the limited, non-transferable right to use the products purchased and the reagents generated from GenCRISPR services and any related material solely for Research Purposes only, not for any Commercial Purposes.