Members of an array of zinc finger proteins specify distinct Hox chromatin boundaries

The partitioning of repressive from actively transcribed chromatin domains in mammalian cells fosters cell-type specific gene expression patterns. During differentiation, this partitioning is reconstructed, reflecting gene expression profiles appropriate to new cellular identities. Yet, the chromatin occupancy of the key chromatin insulator, CTCF, at the developmentally important Hox clusters remains unchanged during differentiation. Thus, dynamic changes in chromatin boundaries must entail other activities, such as the previously identified MAZ insulator. Given its requirement for chromatin loop formation, we examined cohesin-based chromatin occupancy without the known insulators, CTCF and MAZ, and identified a novel family of zinc finger proteins (ZNFs), some of which exhibit tissue-specific expression. Thus far, two of these novel ZNFs facilitate the formation of chromatin boundaries at the Hox clusters that are distinct from each other and from that of MAZ. PATZ1 was critical to the thoracolumbar boundary of Hox clusters in differentiating motor neurons in vitro, skeletal development in vivo, and to looping interactions within the genome. On the other hand, ZNF263 contributed to cervicothoracic boundaries in motor neurons. We propose that these novel insulating activities act in concert with cohesin, alone or combinatorially, with or without CTCF, to implement precise positional identity and cell fate during development. ### Competing Interest Statement D.R. was a cofounder of Constellation Pharmaceuticals and Fulcrum Therapeutics. Currently, D.R has no affiliation with either company. The authors declare that they have no other competing interests.