High-Throughput Metabolomics Service After Gene Knockout

Metabolomics offers a unique perspective on biological states by quantitatively measuring intracellular metabolites. As an integral branch of systems biology, it aims to analyze and interpret the comprehensive set of metabolites within organisms. Utilizing core technologies such as mass spectrometry (MS) and nuclear magnetic resonance (NMR), precise measurements and comparisons of metabolite fluctuations under various conditions are possible, enhancing our understanding of disease mechanisms and metabolic pathways. Advances in technology, particularly the development of high-throughput gene knockout technologies like the CRISPR-Cas9 system, now allow for detailed investigations into the influence of genes on cellular metabolism.

 

High-throughput gene knockout technology enables the simultaneous knockout of hundreds or thousands of genes across diverse cell types and biological models. This technology facilitates the rapid creation of numerous gene knockout cell lines, enabling systematic analyses of gene functions and networks. Integrating metabolomics with MS, researchers can assess the changes in metabolite composition and abundance in cells or tissues following the knockout of specific metabolic genes, examining their effects on metabolic pathways and how genes regulate cellular metabolism at a genome-wide level. For example, the knockout of particular metabolic genes may result in observable alterations in specific cellular metabolites, elucidating their roles in metabolic pathways.

 

MtoZ Biolabs employs a high-resolution metabolomics analysis platform in conjunction with high-throughput gene knockout techniques to offer comprehensive solutions from gene knockout to metabolomics analysis. Our optimized CRISPR/Cas9 system achieves targeted gene modifications, including single and multiple gene knockouts, frameshift mutations, and large sequence deletions in various cell types such as human and mouse cell lines, primary cells, immune cells, and iPS cells. Post-knockout, the metabolites from cultured KO cells are extracted and analyzed using advanced platforms like high-resolution liquid chromatography-mass spectrometry (LC-MS, GC-MS) and NMR, identifying metabolite types and quantifying changes to elucidate the effects of specific knockout genes on metabolic networks.

 

Service Advantages

1. Knockout Efficiency Guarantee

Optimized CRISPR/Cas9 system, over 80% target knockout efficiency at 70%; after gene knockout, we use Sanger sequencing and deep proteomics technology to double-verify the knockout efficiency, double guaranteeing the authenticity of the data.

 

2. Comprehensive Metabolomics Analysis Platform

Equipped with state-of-the-art sample pre-processing instruments and HPLC coupled with high-resolution MS (Orbitrap Exploris 240 & Orbitrap Exploris 480 with FAIMS Pro), our platform supports efficient metabolite extraction, qualitative and quantitative analyses.

 

3. Short Project Cycle

Before conducting gene knockout experiments, we analyze the necessity and expression of genes in different cells to ensure the feasibility of the experiment and reduce the risk of experimental failure; and there is no need for gRNA validation for customers' targets in pre-experiments, greatly shortening the project cycle.

 

4. One-Stop Service

With extensive omics service experience and skilled technical staff, MtoZ Biolabs customizes optimal project plans based on client needs, managing all aspects from sample receipt to project reporting.

 

Case Study

The solute carrier OCTN1 (SLC22A4) functions as an independent transporter, but its key physiological substrates remain unidentified. To explore OCTN1's physiological role, researchers developed OCTN1 gene knockout (octn1 ( -/- )) mice and conducted metabolomics analysis to detect substrates in vivo. The study found a deficiency of the potent natural antioxidant guanine in the blood and organs of octn1 ( -/- ) mice, highlighting OCTN1's crucial role in maintaining guanine levels in vivo and intestines, potentially crucial for preventing intestinal tissue damage and offering a novel approach to diagnosing inflammatory bowel diseases.