How can CRISPR/Cas delivery to the targeted cell be monitored in vivo and in vitro?
I read in one article that in vitro, it can be monitored by adding the antibiotic resistance gene to the plasmid. and after the delivery process by adding the antibiotic to the plat, we can be sure that survived cells took the plasmid which contains both CRISPR components and antibiotic resistance genes!
so is there a specific approach for monitoring the proper delivery in-vivo as I think the last method is not proper for in-vivo study??
Antibiotic resistance genes are for selection or enrichment. You could use fluorescence reporter gene to label the CRISPR expressed cell either in vivo or in vitro. Those reporters could be delivered as fusion parter with CRISPR: eg. Cas9-GFP, or co-expressed with CRISPR: eg. Cas9-2A-GFP.
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