GCaMP6 reporter mice
Posted in Tips
There are a bunch of GCaMP6 mouse lines out now, from Hongkui Zeng (Ai93, Ai95, Ai96 are available), Cris Niell, GENIE (paper), and others. Have you tried any of these? Labrigger has seen data from many of the lines, and they all look good. Tell us what you think in the comments (anonymous comments are always welcome).
…excellent idea. So: why don’t you start?
😉
We are all anxiously waiting to get more information!
I heard good things about the TIGRES for wide-field imaging. Nobody in my filter bubble seems to know a lot about expression levels (!) and patterns of the non-published lines, though.
Are you guys covering this on the SFN pre-meeting?
Cheers,
t
Haven’t done a head-to-head. The Ai lines all look good.
Does anyone have experience of breeding the Ai lines to homozygosity? Any ensuing weirdness in expression and/or animal phenotype?
Cheers,
Mike
Did you get any response on this? Im also interested to know how the GCaMp6 homozygosity will breed.
they all seem to breed very well in our hands…
We bred the Ai95 (Jax 024105) to homozygosity (after crossing with camk2-cre). The animals are fine and expression levels are good.
cheers,
Johannes
Is it possible to order GcAMP mice that already express the indicator in all neurons? (e.g. under Syn1 promoter) From what I see on jackson labs these are all mice that still need to be crossed with a Cre-mouse, but I don’t have the time or breeding facility for that.
@Felix:
We use the Thy1-GCaMP6f mouse line from Jackson and it works beautifully.
Alberto – which Thy1-6f line have you used? (Tg(Thy1-GCaMP6f)GP5.17Dkim by chance?).
We (Pieter Goltstein in our lab) finally tested our first GC6 mice (Cris Niell) under the 2p. Verdict: we were simply blown away. Expression levels and patterns are perfect – good ubiquitous signal, but clearly not overexpressing too much (not a single filled cell). L4 in visual cortex is expressing nicely and we could image down to L6 without problems. Neuropil signal could be an issue but anyways – this is going to change many things for us . I’m rather certain that once we’ve ramped up our breeding this will almost completely replace our virus injections.
We so far did not test the Allen mice but we’ll do so soon.
The only problem is the driver line. Similar to everybody else we use the Mayford CaMK2-tTA line (http://jaxmice.jax.org/strain/007004.html). Sadly, that essentially only expresses in Cortex and a bit of hippocampus. No subcortical expression.
Has anyone tested alternative, preferably ubiquitous tTA lines?
Cheers,
Tobias
Hi Tobias and others,
In essence I am trying to revive this thread 🙂
Have you had a chance to try the Ai lines and compare them to the Cris Niell one? We are thinking of checking some out and would save a lot of effort if someone has done comparisons already. Our application would depend on having a mouse we can cross to different (cre) driver lines and I very much like the tet control as well.
cheers,
Ede
Hi Ede,
sorry: not yet. I will inject some AAV-Cre soon though. I’ll keep you updated.
Best,
T
Hi everyone.
These days, I have trouble identifying my Ai95 reporter mouse with PCR sequencing. The most werid thing is that I have been using standard PCR protocol from Jackson lab (024105), and it has worked perfectly since last month. Up till now, I cannot find the solutions. Can anybody give me some tips? (Maybe I should change anthoer PCR procedure, but I don’t know where to find it)
Thx…
Have the primer stocks been refreshed recently?