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HEK293 GRASP55 and GRASP65 double knockout cell line
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Invented by Dr. Yanzhuang Wang at University Of Michigan

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Catalogue Number 157712
Antigen/Gene or Protein Targets GRASP55 and GRASP65
Parental Line HEK293 cell line
Host Human
Tissue Kidney
Model Knock-Out
Relevance Clustered regularly interspaced short palindromic repeats (CRISPR)/Cas9 technology was used to knock out Golgi reassembly stacking protein of 65 kDa (GRASP65) and/or Golgi reassembly stacking protein of 55 kDa (GRASP55) in HeLa and HEK293 cells. By knocking out GRASPs it was demonstrated that these proteins are essential for Golgi stacking structure formation and accurate posttranslational modifications function in the Golgi. The GRASP knock out cell lines can be used to study the role of GRASPs in Golgi-related biological processes and diseases in which the Golgi is defective (e.g. Alzheimer’s disease, congenital disorders of glycosylation, foot-and-mouth disease, reoxygenation injury and cancer).
Production Details Duplexed sgRNA oligos were digested and ligated into pSpCas9(BB)-2A-GFP(PX458) and pSpCas9(BB)-2A-Puro(PX459)v plasmids to generate GRASP55/65 GFP or Puro plasmids, respectively. CRISPR knock out cells were generated by transfection with GRASP55/65 GFP or Puro plasmids followed by enrichment of GFP-expressing cells by FACS sorting or by selection with 1 µg/ml puromycin, respectively. Individual clones were generated by plating cells at low density and isolating individual colonies. GRASP knock out was confirmed by Western blotting, immunofluorescence, and DNA sequencing.
Research Area Cell Type or Organelle Marker
Notes The HEK293 WT cell line can be provided, as a positive control, if requested with order

References

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References: 1 entry

Bekier ME et al. 2017. Mol Biol Cell. 28(21):2833-2842. PMID: 28814501.


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References: 1 entry

Bekier ME et al. 2017. Mol Biol Cell. 28(21):2833-2842. PMID: 28814501.


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