CRISPR and biochemical screens identify MAZ as a cofactor in CTCF-mediated insulation at Hox clusters

Nat Genet. 2022 Feb;54(2):202-212. doi: 10.1038/s41588-021-01008-5. Epub 2022 Feb 10.

Abstract

CCCTC-binding factor (CTCF) is critical to three-dimensional genome organization. Upon differentiation, CTCF insulates active and repressed genes within Hox gene clusters. We conducted a genome-wide CRISPR knockout (KO) screen to identify genes required for CTCF-boundary activity at the HoxA cluster, complemented by biochemical approaches. Among the candidates, we identified Myc-associated zinc-finger protein (MAZ) as a cofactor in CTCF insulation. MAZ colocalizes with CTCF at chromatin borders and, similar to CTCF, interacts with the cohesin subunit RAD21. MAZ KO disrupts gene expression and local contacts within topologically associating domains. Similar to CTCF motif deletions, MAZ motif deletions lead to derepression of posterior Hox genes immediately after CTCF boundaries upon differentiation, giving rise to homeotic transformations in mouse. Thus, MAZ is a factor contributing to appropriate insulation, gene expression and genomic architecture during development.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • CCCTC-Binding Factor / chemistry
  • CCCTC-Binding Factor / genetics
  • CCCTC-Binding Factor / metabolism*
  • CRISPR-Cas Systems
  • Cell Cycle Proteins / metabolism
  • Cell Differentiation
  • Cell Line
  • Chromatin / metabolism
  • DNA-Binding Proteins / chemistry
  • DNA-Binding Proteins / genetics
  • DNA-Binding Proteins / metabolism*
  • Embryonic Stem Cells / cytology
  • Embryonic Stem Cells / metabolism*
  • Gene Editing
  • Gene Expression
  • Gene Expression Regulation, Developmental
  • Genes, Homeobox*
  • Homeodomain Proteins / genetics*
  • Mice
  • Transcription Factors / chemistry
  • Transcription Factors / genetics
  • Transcription Factors / metabolism*

Substances

  • CCCTC-Binding Factor
  • Cell Cycle Proteins
  • Chromatin
  • Ctcf protein, mouse
  • DNA-Binding Proteins
  • Homeodomain Proteins
  • Rad21 protein, mouse
  • Transcription Factors
  • c-MYC-associated zinc finger protein
  • HoxA protein