All right! This is it! Our story on CRISPR-Cas13d optimization in animal embryos is now out in Developmental Cell! A short thread... 1/9
@Dmc2Cabd @pablodeolavide @CABD_UPO_CSIC @ScienceStowers https://www.cell.com/developmental-cell/fulltext/S1534-5807(20)30587-6

In the Moreno-Mateos lab we are interested in implementing novel CRISPR-Cas tech in vivo and understanding early vertebrate development 2/9
http://www.cabd.es/en-research_groups-128-326-z3-springboard-group-crispr-cas-in-vivo-optimizations-to-understand-early-vertebrate-development-and-human-diseases-summary.html
http://cellcollectives.com/dt_doctors/mamorenomateos/

Early embryonic development is driven by maternal RNAs deposited into the oocyte. However, maternal factors function have remained elusive due to a paucity of techniques for their systematic disruption and assessment. 3/9

Here, we show that #CRISPR-RfxCas13d ( #CasRx) is an effective and precise system to deplete specific mRNA transcripts in #zebrafish embryos. Example: targeting of dnd1 mRNA, a crucial factor for germ cells (green) formation. We targeted other (13!) mRNAs with similar success 4/9

Encouraged by these results, we purified and used Cas13d protein (instead mRNA coding for it). With this, we accelerate the targeting (e.g. nanog degradation in 2 h post injection Protein vs mRNA Cas13) and increase maternal phenotype penetrance 5/9

Finally we tested our optimized CRISPR-Cas13d system in other teleost ( #killifish and #medaka) and #mouse and show that the technology can be used in these organisms as well 6/9

It hass been a fun collaboration (first of many!) with my dear friend Ariel Bazzini @BazziniLab @ScienceStowers led by @gopalkushawah and @Luisherhuertas (1st & 2nd authors respectively) with the contributions of @ItsOverYourHead,D. Monges and @IsmaelMSanchez from our labs 7/9

We also had the luck to collaborate with @proteolaura
from CABD proteomic unit and @JuanRMartinezMo
from @CABD_UPO_CSIC, @JoAbugattas and @malagatrillolab, @Planaria1 and Carter Takacs lab
@UNewHaven 8/9

Thanks to the providers of resources and money for the lab :) Ramon y Cajal program (RyC-2017-23041), grants PGC2018-097260-B-I00 and MDM-2016-0687 ( @Dmc2Cabd) from @AgEInves, the Springboard program from @CABD_UPO_CSIC and Genome Engineer Innovation 2019 Grant from @Synthego 9/9

And that's it! Cheers from Almerimar, Almeria Coast, Andalucia, Spain! :)