A comprehensive HA-tagged PRC1 toolkit enables standardized chromatin profiling and proteomic analysis in Drosophila melanogaster

Authors

Cavalli, G., Fritsch, L., Loubiere, V., Donjon, A., Morales-Sanfrutos, J., Sabido, E., Martinez, A.-M., Schuttengruber, B.

Abstract

Polycomb group (PcG) proteins are evolutionarily conserved epigenetic regulators that maintain transcriptional states during development and are frequently misregulated in disease. Here, we generated a comprehensive collection of endogenously HA-tagged alleles for the core components of Polycomb Repressive Complex 1 (PRC1) and additional chromatin regulators in Drosophila melanogaster using CRISPR/Cas9-mediated genome engineering. We show that endogenous HA tagging preserves protein expression, chromatin localization, and genome-wide binding profiles, enabling direct comparison using a common antibody to analyze distinct PcG subunits. Proteomic analyses recovered known components of canonical and non-canonical PRC1 and PRC2 complexes and identified additional PcG-associated factors. Notably, multiple components of the nuclear pore complex (NPC) were detected as PcG interactors, and genetic analyses confirmed a functional interaction between them. This resource provides a platform for standardized analysis of Polycomb function and chromatin regulation that can easily be extended to other chromatin regulatory complexes and applied across different developmental stages

Preprint server: bioRxiv
The authors list and abstract were imported from bioRxiv on 01 Aug 2026.

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