Tol2 spearheaded by
@koichi_kawakami
has transformed #zebrafish transgenesis. Vector sets to assemble transgenes with (relative!) triggered a prolific jump in tools, including
* https://anatomypubs.onlinelibrary.wiley.com/doi/10.1002/dvdy.21343

* https://anatomypubs.onlinelibrary.wiley.com/doi/10.1002/dvdy.21354

* https://liebertpub.com/doi/10.1089/zeb.2016.1321

What are we adding now?

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1) Minimal promoter vectors🧬🐁

#Enhancers need a minimal promoter to drive transgenes.
To test regulatory elements, we have been using the mouse beta-globin minprom.

We coupled this tiny (129 bp!) promoter in Gateway pME vectors with fluorophores, creERT2, and Gal4.

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The mouse beta-globin minprom plays well with various enhancers we & others have tested over the years. Try it in your constructs!

...and thx @ojtamplin for introducing me to this🙏

Here's a downstream enhancer from mouse Bmp4 (PMID: 19159624) as minprom:EGFP in zebrafish:

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2) mCerulean🔷 & mApple🍎 vectors
#EverythingIsGFP

To expand ease of use of diverse fluorophores, we made Gateway vectors with the blue mCerulean - our current lab favorite!

Here @aburger2009's isolated human HOXP regulatory region driving mCerulean (& mKate2 lens marker!)
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@BlockInTheBack pointed us to the bright, well-behaved red mApple🍎 as alternative to mCherry, dsRED2, et al - and together we built a suite of mApple Gateway vectors.

Here nicely applied by @TheLovelyLab to label pharyngeal endoderm membranes - give it a try!

#AnmAppleADay
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3) p2A as transgene labels

@kwantourage invested into tools to fluorescently label proteins of interest - cloning a beautiful suite of 3' Gateway vectors with viral p2A + diff. fluorophores & localizations.

Fuse your protein of interest with a C-terminally detaching color!
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