Hello all,
Excuse my ignorance but I was looking on addgene for some high fidelity Cas9 nucleases, combined with some selection tools (i.e. GFP and an antibiotic), however most that I found seemed to have no insert site for the sgRNA template. Is it just assumed that you will insert a second plasmid with the sgRNA scaffold?
Any advice would be much appreciated
While you can add the gRNA using a second plasmid there are some vectors that express both Cas9-HF and a gRNA.
For example.
https://www.addgene.org/108295/
https://www.addgene.org/108301/
https://www.addgene.org/108293/
I can't find any that have all conditions checked
https://www.addgene.org/110863/
has Cas9-HF, selection, and GFP but doesn't express a gRNA from the same vector. However there are thousands of CRISPR plasmids on addgene so it is possible that if you try different combinations of search terms you may be able to find one.
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