Get higher Cas9 expression by choosing the best promoter for your cells with SMARTCas9.
Want better gene editing efficiency? Get increased Cas9 expression!
Now SMARTCas9 gives you a CHOICE of promoters for optimal Cas9 expression in your specific cells. To increase your gene editing efficiency, choose the promoter with the highest activity in your particular cell type(s). Higher promoter activity correlates with higher gene editing efficiency!
For a detailed look at the data, see the scientific poster - Cas9 and synthetic crRNA and tracrRNA are effective tools for gene editing in eukaryotic cells.
In addition to being able to optimize Cas9 expression through a choice of promoters, there is a lot more to like about the new SMARTcas9 expression vector!
You have a choice of a fluorescent marker (mKate2 – red channel) or puromycin selection. You can use these makers at least two ways to enrich for an edited population. As mKate2 expression is driven off of the same promoter as Cas9, you can flow sort your transduced cells to select the “red group” with good Cas9 expression. Similarly, you can kill non-transduced cells with puromycin selection to enrich the population for those cells expressing Cas9. Either practice can increase the overall editing efficiency of your cell population!
The plasmid is endotoxin-free and is ready to transfect on the day it arrives, no cloning required! The original S. pyogenes Cas9 coding sequence has been changed to codon usage (same amino acids) that is more efficiently translated in mammalian systems, and this too – along with the optimal promoter for your cells - will help increase intracellular protein levels.
Another way we make the gene editing workflow easier for you is the availability of a fluorescent control plasmid for straightforward transfection optimizations, and as a handy negative control.
Have a look at the technical manual for SMARTCas9 and if you have questions, please contact our scientific support team.
Finally, we provide you with clear, validated protocols and as always the Dharmacon Technical Support Team is here to help you.