I recently used pSpCas9(BB)-2A-Puro(PX462) for knock out the targets in Raw264.7 cells. During experiment from cloning sgRNA to puromycin selection, i thought that EGFP-selection method is better than puromycin-selection, because puromycin-selection method did not suggest complete selection.
So, I`m going to try EGFP-selection methods.
Before I purchase the pSpCas9(BB)-2A-GFP(PX458, from Addgene), Could anyone recommend a CRISPR/Cas9 plasmid(Facs sorting, EGFP)?
Hi Yang Ping, thanks very much, i will choose one more plasmid from that site for failure. thanks!
Dear Lee
I would recommend to use TLCV2 (Addgene #87360), which enables both puromycin and inducible expression of GFP needed for sorting. I also used the lentiCRISPRv2 blast (Addgene #98293) that worked well on RAW cells.
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