Stable Mgat2 Knockout K16 CHO Cell Line
|T6008||1x106 cells / 1.0 ml|
|Description||Using CRISPR technology Mgat2 was silenced from the K16 CHO cells to generate a glycosylation mutant cell line which expresses hybrid type N-glycans. Mgat2 synthesizes the Mannosyl (Alpha-1,6-)-Glycoprotein Beta-1,2-N-Acetylglucosaminyltransferase enzyme (GlcNAcT-II) which plays a key role in the development of mammals in which carbohydrate-dependent interactions facilitate cell signaling, adhesion and migration. Carbohydrate-carbohydrate interaction by the hybrid type N-glycans has been shown to induce faster cell-cell adhesion and cell migration. Further research and discovery on the functions of N-glycans can be assessed with the Stable Mgat2 Knockout K16 CHO Cell Line.|
|Species||Golden Hamster (M. auratus)|
|Species description||Cricetulus griseus (Chinese hamster)|
|Seeding Density||Thaw entire contents into an appropriate T25 flask as specified in the Propagation instructions.|
For Research Use Only
|Mammalian Selection Marker||Puromycin|
|Unit quantity||1x106 cells / 1.0 ml|
|Caution||For Research Use Only|
|Gene Knockdown Method||Mgat2 (UDP-N-acetylglucosamine:-6-D-mannoside 1,2-N-acetylglucosaminyltransferase II) silenced through C residue inserted after the 22nd nucleotide residue causing frameshift mutation|
|Cell Type||Drug Discovery Cells|
Use of PriCoatTMT25 Flasks (G299) or Applied Cell Extracellular Matrix (G422) is required for cell adhesion to the culture vessels. Grow cells inECM-coated culture vessels unless otherwise specified in the Propagation Requirements below.The base medium for this cell line is Prigrow VIII medium available at abm (TM008). To make the complete growth medium, add the following components to the base medium to final concentration of fetal bovine serum (TM999) to a final concentration of 10% and Penicillin/Streptomycin Solution (G255) to a final concentration of 1%. Carbon dioxide (CO2): 5%, Temperature: 37.0°C.
Selection: 4 μg/ml puromycin (G264.)
1) 4 µg/ml of puromycin (G264) was used for clonal selection; 2) Mgat2 silencing was confirmed by DNA sequencing of targeted genomic region; 3) E-cadherin was determined through the use of Western blot method
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|Depositor||Eastern Carolina University|
- Hall, MK et al. "Predominant Expression of Hybrid N-Glycans Has Distinct Cellular Roles Relative to Complex and Oligomannose N-Glycans" Int J Mol Sci. 17(6):E925 (2016). DOI: 10.3390/ijms17060925.