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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:

Icon How_to_use_Tetbow.pdf (719.4 KB)

Plasmids from Article

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