HeLa GRASP55 knockout cell line
- Name Yanzhuang Wang
- Institute University of Michigan
Tool name: HeLa GRASP55 knockout cell line
Tool type: Cell Lines
Tool sub-type: Continuous
Parental cell line: HeLa cell line
Description: 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. Alzheimers disease, congenital disorders of glycosylation, foot-and-mouth disease, reoxygenation injury and cancer).
Research area: Cell Type or Organelle Marker
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 Âľg/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
Additional notes: Cancer Research Technology Limited (trading research tools as CancerTools.org) has been granted a non-exclusive license to the CRISPR-Cas9 technology by ERS Genomics Ltd under the patent rights listed here: https://www.cancertools.org/tool-faqs#hs_cos_wrapper_widget_1649861453796 This license from ERS Genomics Ltd allows CancerTools.org to develop and commercialise CRISPR-Cas9 modified cell lines for research use only. CancerTools.org can provide these modified CRISPR-Cas9 cell lines to companies under a label-use only license
- For Research Use Only
- • Bekier ME et al. 2017. Mol Biol Cell. 28(21):2833-2842. PMID: 28814501.