Josh Tycko


Systematic discovery of protein functions in human cells to understand gene regulation and enable gene therapy


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Josh Tycko


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Department of Neurobiology


Harvard Medical School




Josh Tycko


Systematic discovery of protein functions in human cells to understand gene regulation and enable gene therapy



Department of Neurobiology


Harvard Medical School



High-Throughput Discovery and Characterization of Human Transcriptional Effectors


Journal article


Josh Tycko, Nicole DelRosso, Gaelen T. Hess, Aradhana, A. Banerjee, Aditya Mukund, Mike V. Van, Braeden K. Ego, David Yao, Kaitlyn K. Spees, Peter H. Suzuki, G. Marinov, A. Kundaje, M. Bassik#, Lacramioara Bintu#
Cell, 2020

Semantic Scholar DOI PubMed
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APA
Tycko, J., DelRosso, N., Hess, G. T., Aradhana, Banerjee, A., Mukund, A., … Bintu#, L. (2020). High-Throughput Discovery and Characterization of Human Transcriptional Effectors. Cell.

Chicago/Turabian
Tycko, Josh, Nicole DelRosso, Gaelen T. Hess, Aradhana, A. Banerjee, Aditya Mukund, Mike V. Van, et al. “High-Throughput Discovery and Characterization of Human Transcriptional Effectors.” Cell (2020).

MLA
Tycko, Josh, et al. “High-Throughput Discovery and Characterization of Human Transcriptional Effectors.” Cell, 2020.


Abstract

Thousands of proteins localize to the nucleus; however, it remains unclear which contain transcriptional effectors. Here, we develop HT-recruit - a pooled assay where protein libraries are recruited to a reporter, and their transcriptional effects are measured by sequencing. Using this approach, we measure gene silencing and activation for thousands of domains. We find a relationship between repressor function and evolutionary age for the KRAB domains, discover Homeodomain repressor strength is collinear with Hox genetic organization, and identify activities for several Domains of Unknown Function. Deep mutational scanning of the CRISPRi KRAB maps the co-repressor binding surface and identifies substitutions that improve stability/silencing. By tiling 238 proteins, we find repressors as short as 10 amino acids. Finally, we report new activator domains, including a divergent KRAB. Together, these results provide a resource of 600 human proteins containing effectors and demonstrate a scalable strategy for assigning functions to protein domains.


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