For CRISPR, transfection or transduction?
The CRISPR vectors I bought is lenti vector, so I can either do transfection or transduction.
But which is better??
Hi I'm using lentiviral CRISPR V2 vector from Zhang's lab. https://www.addgene.org/52961/ I'm getting much better results with transduction than standart transfection (Lipofectamine). The puromycin selection after transduction results in almost 100% succes in gene knockout (checked by sequencing). I believe that lentiviral expression, unlike transfection, can provide a more even and more appropriate expression of large proteins, such as CAS9. However, puromycin selection is required to remove uninfected cells.
It really depends on your cells and methods of transfection. If you have an easily transfected cell line, there is no need to go through the effort of making virus for transduction. Test you transfection efficiency with a GFP expressing vector. See how many of your cells are green. If it is a high percentage, you are good to go. Good luck.
There is a lenti focused webinar coming up that might address your concern and help answer any additional questions.
Title: Knockdown, Knockout, Validate: Lentivirus delivers payload in vitro and in vivo Date: Wednesday, May 16, 2018 Duration: 60 minutes Times: 10:00 AM (CST) Speaker: @Christy Hoffmann
What you will learn: Lentivirus biosafety features and best handling practices Flexible customization options to fit your needs Techniques to incorporate lentivirus into your gene studies Insight from a leading Lenti manufacturing expert
Register here: https://www.sigmaaldrich.com/life-science/learning-center/customer-education/lentivirus.html
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