I'm planning to transfect HEK293 cells with lentiCRISPRv2 plasmid containing a gRNA sequence using PEI transfection as non-viral delivery. Would you expect that Cas9 and gRNA expression is affected/interrupted in host cells?
Sanath Kumar Janaka
Expression from Lenticrispr V2 will not be affected by PEI mediated transfection
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Thomas Mühlenberg
It should work alright. I've transected lentiCRISPRv2 transiently by electroporation and you get decent expression and indels. However, puro selection will be necessary. You have to keep in mind that it's a rather large plasmid, which may or may not affect your PEI protocol.
Hope that helps!
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