Pichia pastorise Gene Knock-in Service

Gene knock-in is the use of gene homologous recombination to transfer foreign functional genes (genes that did not exist in the genome or have been inactivated) into cells and genomes. The homologous sequences undergo homologous recombination to insert into the genome, so as to obtain expression in the cell.

There are two types of gene knock-in. One is in-situ knock-in, that is, inserting a new gene at the site of the original gene knock-out, which is the reverse process of gene knock-out. The other is site-directed knock-in, that is, no matter where the knock-out gene is located, the knock-in gene is transposed in the form of a transfer vector under a specific promoter, so the insertion site is certain.

Service introduction

Creative BioMart researchers have established a mature yeast gene editing platform to provide high-quality gene knock-in services for Pichia pastorise and escort the success of your research.

Pichia pastorise genome knock-in technology

• CRISPR / Cas9 method

The CRISPR / Cas9 system is a natural defense system unique to bacteria and archaea, which is used to resist viruses or foreign plasmids.

• Red homologous recombination method

The Red system of Pichia pastorise is used to carry out homologous recombination of foreign DNA to achieve allelic replacement of target genes.

1. Designing the homologous fusion fragment of the target gene

2. Cloning it into the suicide vector

3. Import the suicide vectors into target bacteria by conjugation

4. Screen insertion mutants with antibiotics

5. Screen the surviving strains that undergo the second homologous recombination and lose the suicide plasmid

Customer provides information

• Pichia pastorise strain information
• Knock-in site and replacement sequence information.

Delivery result

• Experiment report, including experimental equipment, reagents, methods, results, conclusions.
• Edited strain

Project cycle

It takes about 35 working days to complete Pichia pastorise gene knock-in service.

Service process

Please let us know the details of your yeast related projects through online consultation. We will reply to you in time and provide you with a comprehensive evaluation report on your project, including the feasibility evaluation of the project and the detailed experimental plan designed by our professionals. As our condition optimization is completed, the result report and other final products will be delivered to you immediately.

Creative BioMart has accumulated many years of experience in the field of yeast research, helping our customers accelerate research in all fields using yeast as a research tool, and improve the overall success rate of the project. Please tell us your project requirements, and we will provide you with a full service from strategy design to final report. If you have any questions, please feel free to contact us.