GRP 78 CRISPR Knockout and Activation

Protocol tips

Upstream tips	Protocol tips	Downstream tips
If transfecting more than one plasmid (i.e., CRISPR/Cas9 KO Plasmid with HDR Plasmid), mix Plasmid DNA at equivalent ratios.	Do not add antibiotics to the Plasmid Transfection Medium

Upstream tips
If transfecting more than one plasmid (i.e., CRISPR/Cas9 KO Plasmid with HDR Plasmid), mix Plasmid DNA at equivalent ratios.
Protocol tips
Do not add antibiotics to the Plasmid Transfection Medium

Publication protocol

GRP78 overexpression
The overexpression of GRP78 was carried out with a GRP78 CRISPR/dCas9 activation plasmid (Santa Cruz, Germany). Cells were transiently transfected with a GRP78 CRISPR/dCas9 activation plasmid (1 μg) or a control scrambled CRISPR activation plasmid using the UltraCruz Transfection Reagent (Santa Cruz, Germany), according to the manufacturer’s instructions. 24 h post-transfection, GRP78 overexpression was verified with Western blot analysis or cells were treated with plumbagin. Samples were analyzed by flow cytometry following Annexin V-PE staining to determine apoptosis induction.



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Manufacturer protocol

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