CRISPR Knock-in in Precision Animal Breeding

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CRISPR Knock-in in Precision Animal Breeding

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Based on its extensive experience in animal genome editing projects and an exceptional molecular biology experimental team, BioVenic has introduced personalized genome knock-in services to customers to address various challenges in transgene knock-in for precision animal breeding research.

Background of CRISPR Knock-in

Using the CRISPR system to achieve gene knock-in in the animal genome circumvents positional effects, gene silencing, and adverse effects on adjacent genes that are typically associated with random insertion methods. After the complex of gRNA and CRISPR nuclease, it causes a DNA double-strand to break at the target site. The donor carrying the knock-in fragment serves as a template for homologous recombination repair (HDR) to insert the fragment into the genomic target site. This process allows for a stable and convenient knock-in of foreign genes or gene fragments.

Fig.1 Schematic Diagram of CRISPR Knock-in System. (BioVenic Original)

Our Services

Knock-in Vector Construction

BioVenic has launched a construction service for knock-in tool vectors, including the construction of CRISPR editing plasmids and donor gene plasmids, to meet the scientific research requirements of customers' exogenous fragments for precise knock-in of genomic sites. While providing high-efficiency editing plasmids, we also offer vector digestion and sequencing verification reports.

Knock-in Protocol Customization

BioVenic specializes in all technologies related to CRISPR knock-in, including the construction of knock-in vectors, cell transfection, and post-knock-in verification, making genome editing no longer complicated. In addition, the advanced CRISPR technology experimental platform and scientific research experts in animal gene editing enable us to provide comprehensive services for your genetic knock-in scientific research project in farmed animals, from project planning to execution, simultaneously.

sgRNA and Homologous Repair Template Design and Synthesis

Knock-in experiments also require the introduction of a donor DNA HDR template designed with homology arms overlapping specific cleavage sites. BioVenic, with the rapid and high-throughput gene synthesis platform, optimizes sequences for heterologous gene knock-in fragments and customizes high-quality sgRNA and HDR templates for target sites, aiming to maximize the editing efficiency of your CRISPR experiments.

Genome Knock-in Modification Validation

BioVenic utilizes the high-throughput sequencing platform to sequence and verify the results of genome knock-in modification, ensuring the smooth advancement of your scientific research projects. We also provide verification services for all genome modifications edited by CRISPR in farmed animals, enabling visualization of gene editing results.

Fig.2 CRISPR Knock-in Methodolog (BioVenic Original)

Applications of Gene Knock-in in Animal Breeding

The application of knock-in editing in precision animal breeding is also quite extensive. Researchers obtain better strains by knocking in to inactivate the target gene or directly knocking in the entire exogenous functional gene. Examples of animal breeding using CRISPR knock-in (KI) include the mouse-derived UCP1 KI pigs with cold resistance improvement, and goats with meat quality improvement by knocking out the MSTN gene and inserting the exogenous fat-1 gene specifically. CRISPR gene knock-in technology creates new breeds of economically valuable farmed animals by breaking species boundaries.

Why Choose Us?

Stable expression of foreign functional genes after insertion.

High-precision seamless knock-in.

No adverse effects on the animal genome.

Relying on extensive experience in scientific research projects, mature technical support, and guidance, BioVenic offers professional and efficient one-stop customized services for your CRISPR knock-in requirements, providing comprehensive and precise knock-in solutions that are tailored to meet your specific breeding research needs. Feel free to contact us for any additional information or pricing inquiries.

References

Zheng, Q., et al. Reconstitution of UCP1 using CRISPR/Cas9 in the white adipose tissue of pigs decreases fat deposition and improves thermogenic capacity. Proceedings of the National Academy of Sciences of the United States of America. 2017,114(45), E9474–E9482.
Zhang, J., et al. CRISPR/Cas9-mediated specific integration of fat-1 at the goat MSTN locus. Federation of European Biochemical Societies. 2018,285: 2828-2839.