Bright multicolor labeling of neuronal circuits with fluorescent proteins and chemical tags.

Sakaguchi R, Leiwe MN, Imai T
Elife. 2018 Nov 20;7. pii: 40350. doi: 10.7554/eLife.40350. PubMed Article

Stochastic multicolor labeling is a powerful solution for discriminating between neurons. Brainbow was developed in 2007 by Joshua Sanes and Jeff Lichtman to use the Cre-loxP system to express one of three fluorescent proteins. When multiple copies of the transgene are introduced, stochastic expression of the three genes will result in the production of dozens of color hues. Now the Imai lab has further developed this technique by including a Tetracycline trans-activator system to boost expression levels of the fluorescent proteins. They named this system Tetbow (Tetracycline transactivator Brainbow).

Download a protocol for how to use TetBow:

How_to_use_Tetbow.pdf 719.4 KB

Plasmids from Article

ID	Plasmid	Purpose
104102	pCAG-tTA	Expresses tTA2 under the CAG promoter in mammalian cells.
104103	pBS-TRE-mTurquoise2-WPRE	Expresses mTurquoise2 under the TRE promoter in mammalian cells.
104104	pBS-TRE-EYFP-WPRE	Expresses EYFP under the TRE promoter in mammalian cells.
104105	pBS-TRE-tdTomato-WPRE	Expresses tdTomato under the TRE promoter in mammalian cells.
104106	pBS-TRE-SNAPf-WPRE	Expresses SNAPf under the TRE promoter in mammalian cells.
104107	pBS-TRE-CLIPf-WPRE	Expresses CLIPf under the TRE promoter in mammalian cells.
104108	pBS-TRE-HaloTag-WPRE	Expresses HaloTag under the TRE promoter in mammalian cells.
104109	pAAV-Syn1-tTA	AAV-mediated expression of tTA under the Syn1 promoter.
104110	pAAV-TRE-mTurquoise2-WPRE	AAV-mediated expression of mTurquoise2 under the TRE promoter.
104111	pAAV-TRE-EYFP-WPRE	AAV-mediated expression of EYFP under the TRE promoter.
104112	pAAV-TRE-tdTomato-WPRE	AAV-mediated expression of tdTomato under the TRE promoter.