I am looking for a human breast cancer cell line with the Flp-in system. The MDA-MD-231 or MCF7 cell lines would be great! Thanks.
Hi eloise,
not yet explore the cas9/talen...friends are starting to use it and they are happy with cas9/crisp easy to set up . the genocoppia kit sounds nice, the experssion casette contain a puromyvine counter selection gene and therefore, it might help getting a stable cell line.
for MCF7...I googled and find out an igem team prepare flp in MCF7..
http://2009.igem.org/Team:Heidelberg/stables
maybe worth getting in touch with them and see if the cell line is still "alive".
personnally, i think in the very next future, it will be useless to keep working with transient transfection or classical stable cell lines (too much overexpression, possible insertional mutagenesis etc etc)...the cas9 community is exchanging idea, experience (see https://groups.google.com/forum/#!forum/crispr)...and the field is going forward ...fast in cell lines, and animal models.
I would advice you to use the aavs targeting system. there is currently a special discount 1600€ . contrary to the flp in system, the aavs targeting is not restricted to a special cell line. we are also thinking to organize a small internal workshop about the new cas9 gene editing. come and we can discuss ...as you are in montpellier
Sorry Alain I cannot help but I am also interested in knock-in and KO in breast cancer cell lines. Genecopoeia offers Talen and CRISPR at reasonable pricing (see Didier's post). They do the constructs and can validate them but are quite reluctant to establish "difficult" cell lines. You will have to provide them with your 231 and MCF7, genotyped and myco-free with the proof of both tests. I've heard Cellectis is good (and it's French so it could be easy for both of you!), but their consumer service is SO BAD that I never heard back from them after sending an inquiry...
Didier: do you recon that TALEN is as good as CAS9?
Hi eloise,
not yet explore the cas9/talen...friends are starting to use it and they are happy with cas9/crisp easy to set up . the genocoppia kit sounds nice, the experssion casette contain a puromyvine counter selection gene and therefore, it might help getting a stable cell line.
for MCF7...I googled and find out an igem team prepare flp in MCF7..
http://2009.igem.org/Team:Heidelberg/stables
maybe worth getting in touch with them and see if the cell line is still "alive".
personnally, i think in the very next future, it will be useless to keep working with transient transfection or classical stable cell lines (too much overexpression, possible insertional mutagenesis etc etc)...the cas9 community is exchanging idea, experience (see https://groups.google.com/forum/#!forum/crispr)...and the field is going forward ...fast in cell lines, and animal models.
Thanks Eloise and Didier...As Didier told me, and mentioned in the Heidelberg team, the problem with the FRT system is the multiple integration sites. I will have a look to the Cas9 system and genocopia.
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