the difference between two systems

all

In <Improved vectors and genome-wide libraries for CRISPR screening>,the article said that 'Both the single- and dual-vector systems mediated efficient knockout of a genomically integrated copy of EGFP in human cells',but in most of articles i have seen,they uesd dual-vector systems for screening. I wonder the difference between the two systems is just to increase viral titer?


Login or Signup to leave a comment
Fuchsia Kelpieover 4 years ago

The 1-vector system will have much lower lentiviral titer than the 2-vector system because the Cas9 is very large, so the molarity of the transfected plasmid when making lentivirus will be lower. Typically for screening we make a large amount of sgRNA library lentivirus to maintain coverage, and if using dual vector-system, you will need to make a lot more lentivirus over all to maintain the same coverage. In addition, if you express the Cas9 first, you will get faster KO when you introduce the sgRNA, which can be helpful depending on your phenotype.

Find your community. Ask questions. Science is better when we troubleshoot together.
Find your community. Ask questions. Science is better when we troubleshoot together.

Have a question?

Contact support@scifind.net or check out our support page.