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.

Catalog Number Product Size AVAILABILITY Price Qty
EMI014
GALE Knock Out Cell Line
1 vial In stock
Price: $499.00
EMI015
GALK1 Knock Out Cell Line
1 vial 3-5 weeks
Price: $499.00
EMI016
GALK2 Knock Out Cell Line
1 vial 3-5 weeks
Price: $499.00
EMI017
GALE/GALK1 Knock Out Cell Line
1 vial In stock
Price: $499.00
EMI018
GALE/GALK2 Knock Out Cell Line
1 vial In stock
Price: $499.00
Specifications
Product Type: Cell Line
Name: GALE KO
GALK1 KO
GALK2 KO
GALE+GALK1 KO
GALE+GALK2 KO
Cell Type: CRISPR modified HEK293T cell lines
Organism: Human
Morphology: Adherent
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
Source: Embryonic kidney
Storage: LN2
Shipped: Dry Ice
Provider
From the laboratory of James M. Termini, PhD, University of Miami.

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. 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.
References
  1. 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.

If you publish research with this product, please let us know so we can cite your paper.

 
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