CRISPR-ChIP reveals selective regulation of H3K79me2 by Menin in MLL leukemia
- Author(s)
- Gilan, O; Talarmain, L; Bell, CC; Neville, D; Knezevic, K; Ferguson, DT; Boudes, M; Chan, YC; Davidovich, C; Lam, EYN; Dawson, MA;
- Journal Title
- Nature Structural & Molecular Biology
- Publication Type
- Research article
- Abstract
- Chromatin regulation involves the selective recruitment of chromatin factors to facilitate DNA repair, replication and transcription. Here we demonstrate the utility of coupling unbiased functional genomics with chromatin immunoprecipitation (CRISPR-ChIP) to identify the factors associated with active chromatin modifications in mammalian cells. Specifically, an integrated reporter containing a cis-regulatory element of interest and a single guide RNA provide a chromatinized template for a direct readout for regulators of histone modifications associated with actively transcribed genes such as H3K4me3 and H3K79me2. With CRISPR-ChIP, we identify all the nonredundant COMPASS complex members required for H3K4me3 and demonstrate that RNA polymerase II is dispensable for the maintenance of H3K4me3. As H3K79me2 has a putative oncogenic function in leukemia cells driven by MLL translocations, using CRISPR-ChIP we reveal a functional partitioning of H3K79 methylation into two distinct regulatory units: an oncogenic DOT1L complex directed by the MLL fusion protein in a Menin-dependent manner and a separate endogenous DOT1L complex, where catalytic activity is directed by MLLT10. Overall, CRISPR-ChIP provides a powerful tool for the unbiased interrogation of the mechanisms underpinning chromatin regulation.
- Publisher
- Springer Nature
- Keywords
- Animals; Humans; *Clustered Regularly Interspaced Short Palindromic Repeats/genetics; Transcription Factors/genetics; Myeloid-Lymphoid Leukemia Protein/genetics; Chromatin; *Leukemia/genetics; Chromatin Immunoprecipitation; Mammals/genetics
- Department(s)
- Laboratory Research; Clinical Haematology
- PubMed ID
- 37679565
- Publisher's Version
- https://doi.org/10.1038/s41594-023-01087-4
- Terms of Use/Rights Notice
- Refer to copyright notice on published article.
Creation Date: 2023-11-30 06:12:31
Last Modified: 2023-11-30 06:21:19