I like Human Hek293 cells because they transfect so well - usually ~95% with Lipofectamine. However I need to work with a Rat cell line to test CRISPR gRNAs in and having a rat cell line that transfects as well would be extremely useful. Thanks in advance.
Back in the last century, we transfected Rat-1a (or Rat-1) fibroblast cell lines with calcium phosphate or cationic liposomes. Worked pretty well.
Thank you to you both. Peter. You don't by chance know someone at NCI that might have the Rat1A cell line or the related Rat2 cell line, do you? It looks like ATCC discontinued the Rat1 and Rat1A lines but they carry the "derived" line Rat 2, which their website claims is highly transfectable. So thank you for suggesting this. If I need to, I'll buy it from ATCC, but I'd rather get it for the cost of shipping if possible. Thanks so much!
I work with rat kidney and we always use COS-7 cells for transfections.
http://www.atcc.org/products/all/CRL-1651.aspx
You can try using NRK cell line, which is not only well established cell line but also easy to transfect. We also use PC12 cell line, which is also easy to grow but difficult to transfect.
Hi Jonathan You can try C6 (rat glioma) that are OK to transfect but not as easy than HEK. I can suggest DreamFect Gold as transfection reagent
http://www.ozbiosciences.com/transfection-dna/19-dreamfect-gold-transfection-reagent.html
Good Luck
While I agree with the earlier suggestion that cos-7 cells are easily transfected, cos-7 is from a green monkey.
INS-1E cells are also adherent rat cells but they are neither easy to grow nor transfectable easily.
Hi Jonathan!
We are starting to try CRISPR in rat as well! Did the B53 line work out for you? It is from the BDIX background, which is different from the actual rat line we will use the CRISPRs in (we will do CRISPR it in animals but test in cell culture first). Is that also the case for you? If so, any experience with how much strain differences matter in practical terms?
Thanks!
Jen
Hi Jennifer, Yes the B53 cells work well. However we have not done much with rat. Most of our work has been in mouse. Using mouse N2A cells has served as a good tool to validate gRNAs that have also worked in vivo, targeting neurons in the mouse brain. I think it is possible that you might see differences between differing cell lines and cell types in vivo. However the risk is relatively low and you don't have much of an alternative. I would test 3-5 gRNAs in cells and use the most efficient for your in vivo work. In cells you should be getting 40-60% editing. I would of course test for gRNA function in vivo too. Regarding strain differences. The main concern might be if you happen to have the bad luck of choosing a gRNA that targets a region that has a SNP and this SNP is different between your cell line and your rat strain and/or the published rat sequencing you are using to design your gRNA. If this is a concern you could sequence the locus. However the risk is low. But not impossible. Good luck!
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