Fut8 Knockout Cell Line-CHO-K1
Cat.No. : CSC-RT2707
Host Cell: CHO-K1 Target Gene: Fut8
Size: 1x10^6 cells/vial, 1mL Validation: Sequencing
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Cell Line Information
Cell Culture Information
Safety and Packaging
| Cat. No. | CSC-RT2707 |
| Cell Line Information | This cell is a stable cell line with a homozygous knockout of Fut8 using CRISPR/Cas9. |
| Target Gene | Fut8 |
| Host Cell | CHO-K1 |
| Size Form | 1 vial (>10^6 cell/vial) |
| Shipping | Dry ice package |
| Storage | Liquid nirtogen |
| Species | Hamster |
| Revival | Rapidly thaw cells in a 37°C water bath. Transfer contents into a tube containing pre-warmed media. Centrifuge cells and seed into a 25 cm2 flask containing pre-warmed media. |
| Mycoplasma | Negative |
| Format | One frozen vial containing millions of cells |
| Storage | Liquid nitrogen |
| Safety Considerations |
The following safety precautions should be observed. 1. Use pipette aids to prevent ingestion and keep aerosols down to a minimum. 2. No eating, drinking or smoking while handling the stable line. 3. Wash hands after handling the stable line and before leaving the lab. 4. Decontaminate work surface with disinfectant or 70% ethanol before and after working with stable cells. 5. All waste should be considered hazardous. 6. Dispose of all liquid waste after each experiment and treat with bleach. |
| Ship | Dry ice |
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Background
Case Study
Applications
The FUT8 gene, formally known as "fucosyltransferase 8," plays a crucial role in the glycosylation process. The gene is located on human chromosome 14 and is responsible for encoding a fucosyltransferase that catalyzes the transfer of fucose to the innermost N-acetylglucosamine (GlcNAc) residue of N-glycans via an α1,6-linkage. The core fucosylation process significantly affects the structural properties and functions of glycoproteins, which are essential for normal cellular functions such as cell signaling, cell adhesion, immune response, and protein stability.
Mutations or dysfunction of the FUT8 gene have been associated with various diseases and conditions. Abnormal glycosylation patterns due to up- or down-regulation of FUT8 activity can lead to abnormal cell signaling and disease progression. For example, reduced FUT8 activity is associated with developmental disorders such as congenital disorder of glycosylation (CDG IIc), while overexpression has been observed in several types of cancer, including liver, colorectal, and lung cancers, where it is thought to play a role in tumor progression and metastasis.
α1,6-fucosyltransferase (encoded by the FUT8 gene) is a key enzyme in mammalian cells that transfers fucose to the innermost GlcNAc residue of the N-glycan via α-1,6 linkages. The presence of core fucose on the Fc region of antibodies can inhibit antibody-dependent cellular cytotoxicity (ADCC) and reduce the therapeutic efficiency of antibodies in vivo. Chinese hamster ovary (CHO) cells are a major production platform for biopharmaceutical manufacturing. Therefore, it is advantageous to generate a FUT8 knockout (FUT8KO) CHO cell line that can be used to produce fully non-fucosylated antibodies.
To understand the role of FUT8 in CHO cell glycosylation, researchers performed a large-scale glycoproteomic study using the FUT8 knockout CHO cell line. A total of 7,127 unique intact glycopeptides (IGPs) containing N-linked glycosyl sites, 928 glycosyl sites, and 442 glycoproteins were identified from FUT8 knockout (FUT8KO) and WT CHO cells. In addition, 28.62% of 442 identified glycoproteins and 26.69% of 928 identified glycosylation sites were significantly changed in FUT8KO CHO compared with wild-type CHO cells. In addition, a decrease in fucosylation content was observed in FUT8KO cells, with the core fucosylated glycans almost disappearing, which was the effect of FUT8 knockout. Meanwhile, a total of 51 glycosylation-related enzymes were quantified in both cell types, 16 of which were significantly altered in FUT8KO cells. These glycoproteomic results revealed that the knockout of FUT8 not only affected the core fucosylation of proteins, but also altered other glycosylation synthesis processes and changed the relative abundance of protein glycosylation.
Figure 1. Changes in fucosylated glycoproteins in FUT8KO CHO cells. (A) Heat map of fucosylated and core-fucosylated glycoproteins in WT and FUT8KO CHO cells. (B) The percentage of core-fucosylated glycoproteins among fucosylated glycoproteins in WT and FUT8KO CHO cells. (C) Changes in fucosylation of protein CD166 (7 glycosites) in CHO cells with FUT8KO. (Yang G, et al., 2021)
The Fut8 gene encodes an enzyme called fucosyltransferase 8, which is involved in the core fucosylation of glycoproteins. The Fut8 gene was knocked out (KO) in the Chinese Hamster Ovary (CHO-K1) cell line in order to study the effects of the absence of this enzyme on glycoprotein function and production, particularly for therapeutic proteins. The following are the main applications of Fut8 KO CHO-K1 cells:
1. Glycoengineering and Biopharmaceutical Production
The Fut8 gene knockout in CHO-K1 cells plays an important role in glycoengineering aimed at producing glycoproteins with improved therapeutic properties. The absence of fucose residues on therapeutic proteins enhances antibody-dependent cellular cytotoxicity (ADCC), making these cells useful for the production of monoclonal antibodies with enhanced efficacy in cancer immunotherapy.
2. Glycosylation Studies
This cell line provides an excellent model for studying the role of fucosylation in protein function. Researchers can analyze the structural and functional effects of glycosylation on various proteins, helping to elucidate the mechanisms underlying diseases associated with glycosylation defects.
3. Cancer Research
Since aberrant fucosylation is associated with cancer progression and metastasis, Fut8 knockout CHO-K1 cells are used to study the role of fucose residues in tumor biology. This can lead to the discovery of new biomarkers and therapeutic targets for cancer treatment.
4. Vaccine Development
The altered glycosylation profiles of proteins expressed in Fut8 knockout CHO-K1 cells can be used for vaccine development. This approach aims to produce more immunogenic glycoproteins that elicit stronger immune responses.
For research use only. Not intended for any clinical use.
