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High-throughput Gene Knockout Metabolomics Analysis

Metabolomics provides a unique perspective on biological states through the quantitative measurement of metabolites within cells. As a key branch of systems biology, it is dedicated to analyzing and interpreting the comprehensive collection of all metabolites within an organism. With core technologies such as mass spectrometry (MS) and nuclear magnetic resonance (NMR), we can accurately measure and compare metabolite changes under different conditions, thereby gaining a deeper understanding of disease mechanisms and metabolic pathways. With technological advancements, particularly the development of high-throughput gene knockout technologies like the CRISPR-Cas9 system, we can now explore at a more detailed level how genes impact cellular metabolic processes.

High-throughput gene knockout technology makes it possible to simultaneously knock out hundreds or even thousands of genes in various cell types and biological models. Utilizing this technology, we can quickly construct a large number of gene knockout cell lines to analyze gene functions and gene networks at a systems level. By combining metabolomics mass spectrometry technology, researchers can analyze changes in the composition and abundance of metabolites in cells or tissues after knocking out specific metabolism-related genes, investigating the impact of particular genes on metabolic pathways and understanding how genes regulate cellular metabolism on a genome-wide level. For example, after knocking out certain metabolism-related genes, changes in specific metabolites within the cell can be observed, thereby revealing these genes' roles in metabolic pathways.

Based on a high-resolution metabolomics analysis platform combined with high-throughput gene knockout technology, Biotage Pekin Biotech (BTP) offers a one-stop solution from gene knockout to metabolomics analysis. Our optimized CRISPR/Cas9 system enables single/multi-gene knockout, frame-shift mutations, and large sequence deletions in human/mouse cell lines, primary cells, immune cells, and iPS cells. After obtaining KO cells, they are cultured, and the metabolites produced by KO cells are extracted and analyzed using high-resolution LC-MS and GC-MS platforms, as well as NMR platforms, to identify changes in metabolite types and levels, revealing the impact of specific knockout genes on metabolic networks. BTP has established seven major testing platforms and has CNAS/ISO9001 dual quality system certification laboratories, aiming to provide you with the highest quality research services. We look forward to collaborating with you, and you are welcome to contact us for more service details!

Service Advantages

1) Knockout Efficiency Guarantee: Our optimized CRISPR/Cas9 system has over 80% target knockout efficiency at 70%. After gene knockout, we use Sanger sequencing and in-depth proteomics technology for dual verification of knockout efficiency, ensuring the authenticity of the data.

2)ComprehensiveMetabolomicsAnalysis Platform: Our metabolomics analysis platform is equipped with comprehensive sample preparation instruments (including cell disruptors, tissue disruptors, tissue grinders, etc.) and ultra-high-performance liquid chromatography high-resolution mass spectrometry (Orbitrap Exploris 240 & Orbitrap Exploris 480 With FAIMS Pro), enabling efficient extraction, qualitative identification, and relative quantification (3D & 4D), as well as absolute quantification analysis of metabolites at the omics level.

3) Short Project Cycle:Before conducting gene knockout experiments, we analyze the necessity and expression levels of genes in different cells to ensure the feasibility of the experiment and reduce the risk of experimental failure. Moreover, there is no need to design pre-experiments for gRNA verification for the customer's targets, significantly shortening the project cycle.

4) One-stop Service:BTP has extensive experience in omics services and professional technical personnel who can customize the optimal project plan according to your needs. You only need to tell us your experimental objective and send samples, and BTP will handle all subsequent project aspects, including sample processing, experimental analysis, data analysis, and project reporting.

Application Cases

The solute carrier OCTN1 (SLC22A4) is an independent transporter whose physiologically important substrates are still undetermined. To study the physiological role of OCTN1, researchers first constructed solute carrierOCTN1gene knockout (octn1^-/-) mice and then conducted metabolomics analysis to identify in vivo substrates. The results showed thatoctn1^-/-mice lacked a natural potent antioxidant—guanosine—in their blood and several organs. This study revealed the critical role of OCTN1 in maintaining guanosine exposure in the body and gut, which may be important for preventing intestinal tissue damage and provides a potential method for diagnosing inflammatory bowel disease.