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An image-based CRISPR screen reveals splicing-mediated control of HP1α condensates

Wong, M. M.-K.; Zhou, S.; Carpenter, C.; Valbuena, R.; Priyadarshini, M.; Arya, A.; Rizvi, A.; Carswell-Crumpton, C.; Wileveau, A.; Lopez-Lopez, G.; Tycko, J.; Yao, D.; Spees, K.; Maynard, J.; Bassik, M. C.; Goodarzi, H.; Sanulli, S.

2026-05-28 genetics
10.1101/2025.09.21.676939 bioRxiv
Show abstract

Heterochromatin Protein 1 (HP1) is a fundamental component of constitutive heterochromatin, forming subnuclear condensates whose regulation and function remain poorly understood. Here, we present an image-based CRISPR screen targeting nuclear factors that identifies splicing as a pivotal pathway regulating HP1 condensates. We discovered that unspliced intronic RNA modulates HP1 condensates by interacting co-transcriptionally with HP1. By modulating the intron content, RNA processing restricts HP1-RNA interactions at chromatin, thus enabling heterochromatin organization. Disruption of HP1 condensates due to enhanced interactions with unspliced RNA leads to loss of heterochromatin and the activation of stress response protective genes. We propose that RNA is a central component of heterochromatin that modulates HP1 condensates, and that RNA processing enzymes act as a surveillance mechanism for condensates by dynamically regulating the network of multi-valent interactions between RNA and chromatin factors. This model underscores the crosstalk between chromatin organization, transcription, and RNA processing, potentially governing broader nuclear functions.

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