1/11: Interested in genetic screening in #organoids? Check out #CRISPRLICHT 💡, our method for loss-of-function screening in cerebral organoids 🧠 out now in @ScienceMagazine. By @christopher_esk and @DLindenhofer. Short thread.
science.sciencemag.org/content/early/…
science.sciencemag.org/content/early/…
2/11: Screening in organoids is difficult because just like animal tissue - but unlike 2D cells - cerebral organoid tissue grows unequally, masking potential loss of function effects.
3/11: To overcome this problem, we developed a #CRISPR screen coupled to massive parallel lineage tracing with exact cell counting using dual barcoding in human cerebral organoids.
4/11: Voila #CRISPR-LICHT: CRISPR-LIneage tracing at Cellular resolution in Heterogenous Tissue.
5/11: We used #CRISPRLICHT to screen through candidate genes for a devastating neurodevelopmental disorder, microcephaly.
6/11: We model-verified 25 new microcephaly genes with an independent primary screen hit validation rate of around 75%
7/11: We looked into one gene in particular: IER3IP1. We show IER3IP1 to regulate ER function with an increase of the UPR in IER3IP1 KO organoids.
8/11: Among the cargoes most affected by perturbed ER processing in IER3IP1 KO organoids are extracellular matrix components.
9/11: Loss of ECM contributes to loss of tissue integrity in IER3IP1 KO organoids, premature progenitor loss and overall fewer cells being generated – microcephaly.
10/11: Overall, we developed #CRISPRLICHT, a method for organoid loss-of-function screening, sensitive enough to discover new biology and define a regulator of ER secretion as a microcephaly gene.
11/11: Thanks to our friends & colleagues at the @viennabiocenter : Simon Haendeler, Roel Wester, Florian Pflug @fgp_phlo_org, Benoit Schroeder, Josh Bagley, Uli Elling @EllingUlrich, Hannes Zuber @johannes_zuber, Arndt von Haeseler. @IMBA_Vienna, @MaxPerutzLabs, @IMPvienna.
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