High-Throughout Gene Knockout Services

    "Knocking out" related mammalian genes is an important method for studying gene function or its potential as a drug target, widely used in research on neurodegenerative diseases such as Alzheimer's disease, cancer, cardiovascular diseases, inflammation, or metabolic diseases. CRISPR/Cas9 is a revolutionary gene-editing technology that can precisely modify the genomic sequence. CRISPR is an immune system in bacteria and archaea that can recognize and cut the genome of invading viruses. By introducing the Cas9 protein and corresponding guide RNA, the CRISPR/Cas9 technology can precisely locate and cut specific DNA sequences, achieving gene knockout, point mutations, and gene insertions. CRISPR/Cas9-mediated gene editing has enormous clinical application potential, and as the technology matures, it is playing an increasingly important role in many fields such as drug target discovery and validation, gene function research, gene therapy, and cancer immunotherapy.

     

    1792491895970172928-1.png

    Neil, C. et al. ACS Synth. 2022.

    Figure 1. CRISPR/Cas9 Gene Knockout Technology

     

    To meet the needs of basic research and pharmaceutical R&D, the scientific research field urgently needs to obtain a series of target genes/full-genome knockout cells. MtoZ Biolabs provides high-throughput gene knockout services using an optimized CRISPR/Cas9 system. Our human whole-genome gene knockout Array library covers the complete human genome, and this library has been fully validated in human primary cells through high-content screening, with over 80% of targets achieving more than 70% knockout efficiency.

     

    MtoZ Biolabs is committed to providing you with the simplest and easiest-to-use high-throughput CRISPR gene knockout services, including virus packaging, plasmid preparation, Cas9 concentration testing, etc., solving key issues such as which cells to choose for knockout, whether knockout is possible, how to perform knockout, and the efficiency of knockout. Single/multiple gene knockouts, frameshift mutations, and large sequence deletions can be performed. You only need to provide the gene name and target cells, MtoZ Biolabs takes care of all subsequent projects, including project evaluation (whether the target gene is expressed in the target cells, whether it is an essential gene), KO experiments, KO cell validation, cryopreservation of KO cells, and delivery of live KO cells.

     

    Service Advantages

    1. Strong Sample Applicability

    Applicable to human and mouse cell lines, primary cells, immune cells, and iPS cells, etc.

     

    2. Analysis of Gene Necessity and Expression Levels

    Before conducting gene knockout experiments, we analyze the necessity and expression levels of genes in different cells based on bioinformatics and proteomics data to ensure the feasibility of the experiment and reduce experimental risks.

     

    3. No Need to DEesign and Validate gRNAs from Scratch

    We have completed the full validation of human whole-genome gRNA, and drug-related targets have also been validated by PCR sequencing, eliminating the need for pre-experimental gRNA validation for customers' targets, significantly shortening the project duration.

     

    4. High Throughput and High Efficiency

    We use an optimized CRISPR/Cas9 system for high-throughput gene knockout, with multiple sgRNA/target gene designs ensuring high knockout efficiency, over 80% target knockout efficiency above 70%, and a high-efficiency KO pool can be provided within a short term (within 10 days), meeting the majority of application scenarios.

     

    5. Double Verification of Knockout Efficiency

    After gene knockout, we use the preferred method in WB experiments for antibody KO verification—Sanger sequencing and deep proteomics technologies to double verify the knockout efficiency, ensuring dual authenticity of the data.

     

    Applications

    1. Gene Therapy

    1792492001284952064-2.png

    Dever, D. P. et al. Nature. 2016.

     

    2. Immune Cell Engineering

    1792492073955463168-3.png

    Roth, T. L, et al. Nature. 2018.

     

    3. Gene Function Research

    1792492142037405696-4.png

    Gordon, D. E, et al. Science. 2020.

     

    4. Drug Target Screening

    1792492227664121856-5.png

    Morgens, D. W, et al. Nat Biotechnol. 2016.

     

    FAQ

    Question 1. What are the Advantages of High Throughput Gene Knockout Technology Compared to Traditional Gene Knockout Techniques?

    High-throughput gene knockout technology has the following advantages over traditional gene knockout techniques such as lentiviral packaging and plasmid methods:

     

    ① Editing Throughput

    1. Traditional gene knockout technology: Low throughput, typically taking 2-3 months to construct dozens of knockout cell lines.
    2. High-throughput gene knockout technology: High throughput, usually capable of constructing several hundred knockout cell lines within 1-2 months.

     

    ② Editing Efficiency

    1. Traditional gene knockout technology: Knockout efficiency is around 30-90%, and is uneven.
    2. High-throughput gene knockout technology: Very high knockout efficiency, reaching 70-100%.

     

    ③ KO Cell Storage

    1. Traditional gene knockout technology: Less than 3000 KO cell reserves.
    2. High-throughput gene knockout technology: Covers the entire human genome, with KO cell reserves as high as 19,000+.
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