A toolkit for programmable transcriptional engineering across eukaryotic kingdoms
This article has been Reviewed by the following groups
Discuss this preprint
Start a discussion What are Sciety discussions?Listed in
- Evaluated articles (Arcadia Science)
Abstract
Chromatin is essential for eukaryotic life. Tens of thousands of chromatin regulator (CR) proteins exist in eukaryotic genomes that are predicted to modulate chromatin states; however, their molecular functions remain largely untested experimentally. Here, we construct a library of over 300 full-length CRs from humans, plants, yeast, protozoa, and virus, each fused to DNA-binding domains, and test their direct effect on transcriptional repression and activation in plants and human cells. We discover CRs with cross-kingdom functionality when transferred across eukaryotes, including CRs that outperform existing tools for programmable transcriptional repression and activation in plants and human cells. Using pooled CRISPR screens, we demonstrate a suite of CRISPR repressors that titrate gene expression at intermediate levels. Finally, we identify RCOR1 and MTA2 as universal eukaryotic repressors that retain repressive activity in plants, yeast, and human cells. Our toolkit advances synthetic eukaryotic engineering and expands our understanding of CR functionality across eukaryotes.
Article activity feed
-
DISCUSSION
I see the appeal of identifying evolutionarily agnostic regulatory activity and appreciate the effort to tackle this goal. I was hoping you could speak toward the magnitude of the effects observed in the initial screens.
In the plant and human screens, the largest log fold changes reported appear relatively modest. Do you think this reflects limitations of the screening platforms themselves, the size or composition of the candidate CR library (e.g., that stronger regulators may not have been captured), or might it suggest that CRs lose some regulatory potency when moved outside of their native evolutionary or chromatin context?
-
