Please enjoy our new #preprint, "High-throughput discovery and characterization of human transcriptional effectors"! 😀
We tested thousands of protein domains and mutants for the ability to repress/activate #GeneExpression and ID effectors in 600 proteins
biorxiv.org/content/10.110…
We tested thousands of protein domains and mutants for the ability to repress/activate #GeneExpression and ID effectors in 600 proteins
biorxiv.org/content/10.110…
For many human #TranscriptionFactors & #chromatin regulators, we know more about *where* they bind than *how* they regulate expression
To find effector domains in an unbiased way, we tested a library of the majority of nuclear protein domains with our new method, HT-recruit
To find effector domains in an unbiased way, we tested a library of the majority of nuclear protein domains with our new method, HT-recruit
We find hundreds of human #repressor domains, including several domains of unknown function
Some highlights from testing entire domain families:
In #KRAB, the largest family of repressors, we observe that young domains are repressors while ancient domains are not
Among #HOX homeodomains, we find collinearity between repression strength and Hox genetic organization
In #KRAB, the largest family of repressors, we observe that young domains are repressors while ancient domains are not
Among #HOX homeodomains, we find collinearity between repression strength and Hox genetic organization
We also used HT-recruit to discover activator domains. We find domains of unknown function that work as activators and, surprisingly, the strongest activator was one of the ancient, non-repressive, KRAB domains.
Beyond discovering new effectors, we were also motivated to better understand/optimize a well-known effector used in #SynBio
We did a #DeepMutationalScan of the KRAB used in #CRISPRi to map its co-repressor binding surface and discover KRAB variants with enhanced activity
We did a #DeepMutationalScan of the KRAB used in #CRISPRi to map its co-repressor binding surface and discover KRAB variants with enhanced activity
To go beyond screening Pfam domains, we did unbiased tiling of 238 nuclear proteins and discovered repressors in the unannotated regions of proteins.
For example, in MGA, a repressive TF, we find two repressors that are 10AA and 30AA long
For example, in MGA, a repressive TF, we find two repressors that are 10AA and 30AA long
On a technical note, we made these screens easier by engineering a synthetic surface marker reporter and using magnetic cell separation 🧲instead of cell sorting. #HighThroughput #TechDev
This project is a really fun collaboration between my co-advisors Lacra @BintuLab and Mike Bassik. I'm so thankful for them and labmates including Nicole DelRosso, Aradhana, Gaelen and our collaborators @Abhi2718281828 & @anshulkundaje ! @StanfordBiosci
Please let me know if you have a favorite nuclear protein and we can look into its effector functions! 🔍
Our paper on HT-recruit to discover human #transcription effectors just got published! authors.elsevier.com/c/1cFBAL7PXcLyN
Science 🧬 in the thread 🧵 above 👆
Thank you to the reviewers, editors, and staff @CellCellPress !
#GeneRegulation #EpigenomeEditing
Science 🧬 in the thread 🧵 above 👆
Thank you to the reviewers, editors, and staff @CellCellPress !
#GeneRegulation #EpigenomeEditing
This paper is the result of a fantastic and fun collaboration between my coadvisors @BintuLab and Mike Bassik. Please reach out to any of us if you have questions!
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