Gene Knock-in Services with CRISPR/Cas9 Technology

CRISPR/Cas9 technology is an emerging method which can be used to improve crop properties such as high yields and nutrients, resistance to disease, pests, or drought tolerance. Based on the homology-directed repair (HDR) pathway of the DSB, the CRISPR/Cas9 system can be used for precise knock-in genetic manipulation. Lifeasible has a wealth of successful gene editing cases and can provide a full set of plant genome editing solutions, including gene knock-in and many other gene editing technologies.

Figure 1. The applications of CRISPR/Cas9 in plant. (Li H, et al. 2017)

CRISPR/Cas9 technology greatly reduces the difficulty of the technique of targeted insertion or replacement (Knock-in) of a sequence on a plant chromosome. When Cas9/gRNA cleaves the target site, the gene fragment located on the donor DNA can be integrated into the DSB site by HDR recombination if the cell has a DNA donor fragment homologous to the target site sequence. Lifeasible has developed a proven platform for gene knock in in many plant species.

• Service Strategy
• Using a proprietary vector for DNA donors, the efficiency of knock-in was enhanced by increasing the copy number of donor DNA.
• Cas9 and a pair of gRNAs are used to "cut" a sequence from the DNA donor and then "paste" it into the target site of the plant genome. This strategy utilizes the NHEJ pathway that dominates DSB repair.

Figure 2. Outline of the sequential transformation strategy for gene targeting and screening procedure. (Miki D, et al. 2018)

• Service Flow
• Design and vector construction of sgRNA targeting target sequences
• DNA donor vector construction
• Target plant transformation
• Screening and identification of positive plants
• Positive seedlings hardening

• Technical Advantages
• The latest Nickase doubles gRNA technology, significantly reduce the off-target rate.
• No genus restriction, suitable for targeted knock-in of various plants.
• The short service cycle.
• Free choice of gene insertion, cloning vectors, target plants, and promoters and selective markers.

• Why Choose Lifeasible

Lifeasible provides one-stop technical services from bioinformatics analysis, gene editing molecular experiments, and plant experiments. Our optimized CRISPR/Cas9 system is suitable for most gene target sites and plant species. Lifeasible has a technical team with rich experience in CRISPR gene editing and currently can realize services including gRNA design, CRISPR/Cas9 vector construction, Donor vector construction, transformation, positive seedling identification, seedling hardening and seed harvest, etc. We provide the most cost-effective services, and efficiency and professionalism are the most commented by our customers about Lifeasible.

• How to Place an Order

Lifeasible offers complete, professional plant gene knock-in solutions, as well as customized experimental protocols based on your project requirements and plant sample characteristics. For more information, please contact Lifeasible.