Gene Knockin

In gene editing, gene knockin refers to a genetic engineering method, which involves the one-for-one substitution of DNA sequence information in a genetic locus, or the insertion of sequence information not found within the locus. BioVenic provides the most comprehensive and effective solution for gene knockin workflow. We are committed to helping customers solve all problems encountered in the process of gene tapping, to promote the process of animal breeding. Each of our deliverables has undergone strict quality inspection and testing to ensure the reliability and accuracy of the results.

Gene Knockin

• Point Mutation

As a gene mutation, a point mutation is a change in a single nucleotide base from a single base pair in the genomic DNA or RNA sequence. Broadly defined point mutations can be base substitutions, single base insertions or base deletions; narrowly defined point mutations are called single base substitutions. Base substitutions are divided into two categories: conversions and reversals.

• Gene Insertion, Deletion and Replacement

Gene insertion is the addition of one or more genes to a DNA sequence and is a technique performed using plasmid DNA or integrating viral vectors on conventional methods. As a type of mutation, gene deletion refers to the complete inactivation of the target gene through the loss of genetic material. In contrast, gene replacement is a technique that replaces an endogenous gene at the original locus in the genome, which can delete a gene, delete an exon, add a gene, and introduce a point mutation, among others. Gene replacement can be permanent or conditional.

• Fluorescent Tagging

Fluorescent reporter gene knockin enables targeted fluorescent tagging tasks and is widely used in biomedical fields. Fluorescent tagging has revolutionized biomedical research because it is easy to use for protein labeling and does not require fixation or permeation and is widely used for live cell imaging in various models. Among them, GFP is widely used to study gene expression, characterize protein localization/co-localization, and reveal cellular signaling pathways. DNA cleavage genes ZFNs, TALENs and CRISPR/Cas9 are fluorescently labeled by knocking in fluorescent reporter motifs using gene editing techniques.

Techniques for Gene Knockin

• ZFNs for Gene Knockin

ZFNs are gene editing techniques that fuse the nuclease domain of the FokI enzyme to three (or more) zinc finger (ZF) arrays by binding DNA cleavers to specific DNA recognition sites. ZFNs disrupt the double-stranded structure of DNA when two zinc finger modules with DNA binding sites dock with each other to form a homologous complex with the intervening FokI enzyme, leading to random insertional mutations.

• TALENs for Gene Knockin

TALEs are designed to fuse with the DNA cleavage structural domain of FokI nucleases and are known as TALENs gene editing tools. Each repeat of TALEs contains 33-35 amino acids and recognizes a single nucleotide. TALEs recognize DNA in a modular fashion.

• CRISPR/Cas9 for Gene Knockin

The CRISPR/Cas9 technology uses a specific RNA sequence called a guide RNA to bind to another DNA target sequence (target DNA), and then Cas9 splits where the binding occurs. This makes the CRISPR/Cas9 system the superior gene editing tool because it improves the efficiency and accuracy of gene editing. IF the homologue-directed repair (HDR) happed, with a DNA donor, CRISPR/Cas genome editing can be used to replace an undesirable gene or overexpress (knockin) and an individual gene.

Fig.1 Application of CRISPR/Cas genome editing in gene functional study. (Zhang, 2020)

Our Services

Thanks to our powerful gene editing platform, BioVenic is proud to provide precise and efficient gene knockin services that is committed to ethical and regulatory compliance to achieve animal breeding goals and accelerate the animal breeding process. We have a team of scientists specializing in animal gene editing, who works closely with you and understands your research goals to develop a customized gene knockin strategy that meets your specific needs. Our gene knockin methods including but not limited to.

• ZFNs
• TALENs
• CRISPR/Cas9

Following the knockin process, BioVenic is dedicated to offer a varitey of services for animal gene modification, including gene transfection service, production of genomic modified animals, and gene editing validation services.

Want to Learn More?

As a rapidly growing biology company, BioVenic is committed to helping our customers meet the growing and evolving needs of the animal breeding process. We have a wide range of proven gene editing technologies to help our customers achieve gene knockin. Our proven and well-established technology platform will meet specific animal breeding goals with a commitment to ensuring better animal welfare and a focus on sustainable development. If you are interested in our services or have any further questions, please feel free to contact us.

Reference

1. Zhang D.; et al. CRISPR/Cas: A powerful tool for gene function study and crop improvement. J Adv Res. 2020, 29: 207-221.