CRISPR/Cas9-Based Strain Engineering

Based on our advanced technology and rich experience, BOC Sciences can provide strain development and strain modification/optimization services to enhance their metabolic capabilities. Based on the powerful tool of CRISPR/Cas9, Our strain development and optimization services cover various types of microorganisms, including fungi, bacteria, algae and more.

Fig.1 Development of CRISPR/Cas9-based genome editing tools for polyploid yeast. (Gu et al., 2023)

CRISPR/Cas9 Engineering Services

BOC Sciences utilizes CRISPR/Cas9 engineering as a powerful and precise tool to develop and optimize strains, and to produce strains at scale to meet customer needs for strain customization and to accelerate the project development process.

• Fungal strain development

Fungi include many important biological resources, such as edible mushrooms and enzyme-producing bacteria. Through CRISPR/Cas9 engineering, the genomes of fungi can be edited to change their characteristics such as adaptability and product synthesis.

• Bacterial strain development

Bacterial strain development by CRISPR/Cas9 technology is the use of CRISPR/Cas9 technology to edit and modify the genome of bacteria for strain optimization and engineering for specific purposes. The strain improvement methods include gene knockout, gene insertion, gene modification, and other operations.

• Scale-up Production of Strains

After the optimization of strains, the evolutionary process of strains can be accelerated, so that excellent strains with desired traits can be obtained quickly. Adequate safety assessment and quality control must be performed when using CRISPR/Cas9 technology in the process of large-scale production.

CRISPR/Cas9 Microbial Genome Editing

• Targeted Gene Editing

CRISPR/Cas9 targeted gene editing technology is highly efficient, precise, and flexible. CRISPR/Cas9 can also be used to target the deletion or inactivation of unwanted genes in strains to reduce the accumulation of unwanted metabolic pathways or byproducts. With CRISPR/Cas9-targeted gene editing, gene knockout, gene insertion, point mutation, gene repair and other operations can be realized.

• Genomic Regulation

Precise regulation of genes is required by combining Cas9 proteins with specific regulators, such as activators or repressors. In CRISPR/Cas9 gene regulation, a variant known as dCas9 (dead Cas9) is commonly used, which is an inactive form of the Cas9 protein, i.e., it loses DNA cleavage function. dCas9 proteins can still bind to the DNA sequence of the target gene, but do not cause DNA breaks. With CRISPR/Cas9 gene regulation, we can precisely manipulate gene expression levels to study gene function, investigate gene regulatory networks.

• Genome Rearrangement

CRISPR/Cas9 gene rearrangement is a method that utilizes CRISPR/Cas9 technology to alter the arrangement or structure of genes in the genome. Through the guidance and cleavage functions of the CRISPR/Cas9 system, gene rearrangement, transposition or adjustment can be realized. By rearranging genes, we can explore the functions and regulatory mechanisms of genes in different arrangements or structures. In addition, CRISPR/Cas9 gene rearrangement technology can be used to change the position of specific genes in the genome, thereby adjusting gene expression and interactions.

CRISPR/Cas9 Function in Microorganisms

CRISPR/Cas9 gene editing is a technique for specific DNA modification of target genes and is at the forefront of gene editing technology.

• Rapid Preparation of Cell Models

By using Cas9-mediated genome editing, transgenic models can be generated more rapidly, expanding biology research beyond traditional lab models.

• Functional Genomics Screening

The CRISPR/Cas9 technology is being used to carry out genome-wide functional screenings that enable researchers to discover and identify the genes that contribute to the aggregation of desired phenotypes with extreme sensitivity and precision.

• In Vivo Imaging of Intracellular Genomes

CRISPR/Cas9-based fluorescently labeled Cas9 tags at specific DNA sites have replaced DNA-FISH technology to develop into a powerful live-cell imaging technique that will be used to study complex chromosomal structures and nuclear organization.

Reference

1. Gu L, et al. Development of CRISPR/Cas9-Based Genome Editing Tools for Polyploid Yeast Cyberlindnera jadinii and Its Application in Engineering Heterologous Steroid-Producing Strains[J]. ACS Synthetic Biology, 2023, 12(10): 2947-2960.