The ability to quantitatively study mRNA translation using SunTag imaging is transforming our understanding of the translation process. Here, we expand the SunTag method to study new aspects of transl...

The ability to quantitatively study mRNA translation using SunTag imaging is transforming our understanding of the translation process. Here, we expand the SunTag method to study new aspects of transl...

DNA methyl groups are selectively removed at target loci using inactive Cas9 fused to a SunTag-based peptide repeat that recruits the enzymatic activity.

Cells communicate via extracellular vesicles (EVs) containing functional RNAs, proteins and lipids. Knowledge on the fate of internalized EVs, especially their capacity to fuse with target cell membra...

CRISPR activation (CRISPRa) is a powerful tool for endogenous gene activation, yet the mechanisms underlying its optimal transcriptional activation remain unclear. By monitoring real-time transcriptional bursts, we find that CRISPRa modulates both burst duration and amplitude. Our quantitative imaging reveals that CRISPR-SunTag activators, with three tandem VP64-p65-Rta (VPR), form liquid-like transcriptional condensates and exhibit high activation potency. Although visible CRISPRa condensates are associated with some RNA bursts, the overall levels of phase separation do not correlate with transcriptional bursting or activation strength in individual cells. When the number of SunTag scaffolds is increased to 10 or more, solid-like condensates form, sequestering co-activators such as p300 and MED1. These condensates display low dynamicity and liquidity, resulting in ineffective gene activation. Overall, our studies characterize various phase-separated CRISPRa systems for gene activation

Highlighted Article: SunTag imaging in the Drosophila embryo can be exploited to compare translation of maternal and zygotic transcripts, investigate ribosome pausing and study translation of secreted...