O-linked Glycosylation Knock Out Cell Lines
These five cell lines were engineered to be deficient in O-linked glycosylation by knocking out key enzymes of the O-glycoslyation pathway using Crispr/Cas9. These include UDP-galactose-4- epimerase (GALE), galactokinase 1 (GALK1) and galactokinase 2 (GALK2).
Highlights:
- Cell lines were validated using mass spec, sequencing, and western blot analysis
- GALE KO- Deficient in O-glycans in normal media but media + GalNac and Galactose, cells restore O-glycosylation to normal levels
- GALE+GALK1 KO - Devoid of O-linked glycan as well as only contain high-mannose N-glycans
- GALE-GALK2 KO - Completely devoid of O-glycans and can not be restored no matter exposure to galactose or GalNac
O-linked glycosylation is the attachment of a sugar molecule to an oxygen atom in an amino acid residue in a protein. O-linked glycosylation is a form of glycosylation that occurs in the Golgi apparatus in eukaryotes. These key enzymes are critical for glycan salvage and synthesis of UDP-galactose & UDP-GalNac. These cell lines are ideal for many studies where one wishes to investigate whether O-glycans are important or play a role in their research.
From the laboratory of James M. Termini, PhD, University of Miami.
These five cell lines were engineered to be deficient in O-linked glycosylation by knocking out key enzymes of the O-glycoslyation pathway using Crispr/Cas9. These include UDP-galactose-4- epimerase (GALE), galactokinase 1 (GALK1) and galactokinase 2 (GALK2).
Highlights:
- Cell lines were validated using mass spec, sequencing, and western blot analysis
- GALE KO- Deficient in O-glycans in normal media but media + GalNac and Galactose, cells restore O-glycosylation to normal levels
- GALE+GALK1 KO - Devoid of O-linked glycan as well as only contain high-mannose N-glycans
- GALE-GALK2 KO - Completely devoid of O-glycans and can not be restored no matter exposure to galactose or GalNac
O-linked glycosylation is the attachment of a sugar molecule to an oxygen atom in an amino acid residue in a protein. O-linked glycosylation is a form of glycosylation that occurs in the Golgi apparatus in eukaryotes. These key enzymes are critical for glycan salvage and synthesis of UDP-galactose & UDP-GalNac. These cell lines are ideal for many studies where one wishes to investigate whether O-glycans are important or play a role in their research.
From the laboratory of James M. Termini, PhD, University of Miami.
| Catalog Number | Product | DataSheet | Size | AVAILABILITY | Price | Qty |
|---|
| Product Type: | Cell Line |
| Cell Type: | CRISPR modified HEK293T cell lines |
| Accession ID: | Q14376, P51570 |
| Organism: | Human |
| Morphology: | Adherent |
| Source: | Embryonic kidney |
| Biosafety Level: | BSL1 |
| Subculturing: | Split 1:10 to 1:20 when cells reach 80% confluence |
| Growth Conditions: | DMEM + 10% FBS + Pen/Strep |
| Cryopreservation: | 90% FBS + 10% DMSO |
| Comments: | The GALE KO cell line is deficient in O-glycans when grown in normal media however, when media is supplemented with GalNac and Galactose, these cells restore O-glycosylation to normal levels. All cell lines were validated using mass spec, sequencing, and western blot analysis. See included publication. GALE+GALK1 KO were found by western blot to be devoid of O-linked glycan as well as only contain high-mannose N-glycans. GALE-GALK2 KO are very useful because they are completely devoid of O-glycans and can not be restored no matter exposure to galactose or GalNac. All important data is in publication. |
| Storage: | LN2 |
| Shipped: | Dry Ice |
- Termini JM, Silver ZA, Connor B, Antonopoulos A, Haslam SM, Dell A, Desrosiers RC. HEK293T cell lines defective for O-linked glycosylation. PLoS One. 2017 Jun 27;12(6):e0179949. doi: 10.1371/journal.pone.0179949. eCollection 2017. PubMed PMID: 28654657; PubMed Central PMCID: PMC5487050.
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