GCaMP3 is now officially old skool. GCaMP5 is available at addgene.
Plasmid 31788: pCMV-GCaMP5G, a.k.a. GCaMP3-T302L R303P D380Y
Dimmer baseline F, higher dF/F, picks up significantly more activity in vivo cortex. Still not holy grail level, but getting closer.
How does it compare to the Campbell group’s GECOs?
Nobody knows, not even Rob. I believe his collaborators are trying to get data on action potential detection, but I’m not aware of any actual results on that front yet. The GECOs look potentially great for multicolor imaging, but until someone does in utero or virus injections with them there is no way to know.
Many, many sensors that look great in vitro fail miserably in vivo.
Here it is:
http://www.frontiersin.org/cellular_neuroscience/10.3389/fncel.2012.00041/abstract
Unfortunately, it seems that GECOs do not show any improved performance in mammalian neurons, when characterized with electrophysiology in acute brain slices.
Great news. Any chance it’s available in an AAV plasmid (ie something akin to AAV-Syn-GCamP3 and ready to package)? Or better yet, are viral stocks available through UPenn or elsewhere? Thanks for the update.
Available via U-Penn:
V2340TI – AAV2/1.Syn.GCaMP5.WPRE.SV40 (p2401)
Cheers, Christian
Thanks!
[x] ordered
…and now the FLEXed virus, please!
Sadly, this does not appear to be in the near future for U Penn Vector Core.
Update on that: U Penn have completely revamped their catalogue. FLEXed versions of GCaMP5 with hSyn or CAG promoters, different AAV serotypes, etc. are now available. Sometimes it’s nice to be wrong.
Is the comparison w/ GCaMP3 ever going to be published, or is the world just moving onto 5?
Ask the reviewers/editor. Its a shame how difficult it is to get better sensors published in decent journals, given their broad utility and extremely high citation rate.
Also, membrane targeted version now on Addgene – http://www.addgene.org/34924/
Are the GCaMP5 spectra essentially the same as for GCaMP3? Thanks!
Yes.
Thanks a lot for your info. Do you know if I could get .Syn.GCaMP5.WPRE.SV40 construct from Upenn?