Comprehensive CRISPR Gene Editing Solutions to Power Your Research: KO Cell Lines, CRISPR Screen & More

Gene knockout (KO) is typically permanent, especially when using genome-editing techniques like CRISPR-Cas9. Here's why:

• Mechanism: In gene knockout, a DNA double-strand break is introduced at a specific location in the gene. This break is repaired by the cell’s own repair mechanisms, which often result in insertions or deletions (indels) and introduce an early stop codon,that disrupt the gene's coding sequence, causing it to lose its function. These mutations are passed on to daughter cells when they divide.
• Permanent in Cell Lines: If you're generating a knockout in a cell line, the knockout is usually stable over time, as the altered DNA will be present in every cell, and the mutation is passed down with cell division. You can even create clonal populations where all cells carry the same knockout mutation.
• Permanent in Organisms: If you're creating a knockout in an organism (such as mice or other model organisms), the gene disruption will be passed on through heredity if the knockout is done in germline cells (e.g., sperm or eggs), leading to permanent knockouts in offspring. In some cases, you might generate heterozygous knockouts first, and then breed them to produce homozygous knockouts for more complete disruption.

The purpose of a gene knockout (KO) is to study the function of a specific gene by completely eliminating its expression or disrupting its function. By knocking out a gene, researchers can observe how the organism or cell behaves in the absence of that gene's product (usually the protein it encodes). This helps answer key biological questions and has a wide range of applications

• Molecular Confirmation Methods:
  • PCR and Gel Electrophoresis
  • Sequencing
  • Southern Blotting
  • Western Blotting (Protein Level Confirmation)
  • qRT-PCR (Quantitative Reverse Transcription PCR)
• Functional Assays:
  • Phenotypic Analysis
  • Reporter Genes
  • Flow Cytometry (for Knockout Cells)
• Knockout Mouse/Model Organism Specific Verification:
  • Breeding and Genotyping (for Mice or Other Organisms)
  • Histological Analysis

Ubigene performs genotypic validation of knockout (KO) cell lines using PCR amplification combined with Sanger sequencing to confirm successful gene disruption at the DNA level. For customers requiring additional verification, customized assays are available as supplementary services. These may include Western blotting or qPCR, depending on the target gene and specific experimental requirements.

Ubigene implements stringent quality control measures for every knockout (KO) cell line. All in-stock KO cell lines undergo comprehensive quality assurance testing prior to shipment. Each cell line is confirmed to be free from Mycoplasma contamination and authenticated by short tandem repeat (STR) profiling to verify its genetic identity and ensure authenticity.

Upon delivery, Ubigene provides a complete documentation package that includes the Mycoplasma testing report, STR authentication report, and detailed cell culture instructions. All KO cell lines are guaranteed to be contamination-free, viable, and ready for downstream experimental applications. In addition, Ubigene offers complimentary technical support and culture guidance to help customers achieve optimal cell performance and experimental outcomes.

Ubigene provides two vials of a validated homozygous knockout (KO) single-cell clone (1 × 10⁶ cells per vial) as the standard deliverable. Each KO clone is isolated from a single cell to ensure genetic homogeneity and stable knockout efficiency.

However, if a KO cell pool is more suitable for your experimental design, Ubigene can also customize and deliver a KO cell pool upon request.

Upon receipt, Ubigene’s KO cells are delivered as cryopreserved vials. To maintain cell viability and ensure optimal experimental performance, please follow the storage and handling guidelines below:

• Immediate storage: Upon arrival, promptly transfer the vials into liquid nitrogen (LN₂) for long-term storage. This ensures maximal preservation of cell viability and genetic stability.
• Short-term storage: If liquid nitrogen storage is not immediately available, the vials may be temporarily stored at −80 °C for up to several days. Avoid prolonged storage at −80 °C and prevent any freeze–thaw cycles, as they may significantly reduce cell viability.
• Thawing and recovery: Ubigene provides a detailed Cell Use Instruction with each shipment. Please follow the recommended thawing and culture procedures carefully to ensure optimal cell recovery and growth.

Note: Always handle cryopreserved cells under aseptic conditions and minimize exposure to ambient temperature during transfer. Proper storage and handling are critical to maintaining cell viability, genetic integrity, and experimental reproducibility.

Don't worry — Ubigene provides 24/7 technical support to assist you with any challenges you may encounter during cell recovery or subsequent experiments. If you experience any issues, simply leave us a message or contact our technical support team. Our specialists will review your case and provide professional guidance within 24 hours of receiving your inquiry.

Ubigene is committed to ensuring the success of your experiments. From cell thawing and culture optimization to troubleshooting and data interpretation, our technical experts are here to provide timely, personalized assistance every step of the way.