Animal chromosomes are partitioned into contact domains. Pathogenic domain disruptions can result from chromosomal rearrangements or perturbation of architectural factors. However, such broad-scale alterations are insufficient to define the minimal requirements for domain formation. Moreover, to what extent domains can be engineered is just beginning to be explored. In an attempt to create contact domains, we inserted a 2-kb DNA sequence underlying a tissue-invariant domain boundary—containing a CTCF-binding site (CBS) and a transcription start site (TSS)—into 16 ectopic loci across 11 chromosomes, and characterized its architectural impact. Depending on local constraints, this fragment variably formed new domains, partitioned existing ones, altered compartmentalization and initiated contacts reflecting chromatin loop extrusion. Deletions of the CBS or the TSS individually or in combination within inserts revealed its distinct contributions to genome folding. Altogether, short DNA insertions can suffice to shape the spatial genome in a manner influenced by chromatin context.

动物染色体被划分为接触域。致病域破坏可能是由染色体重排或结构因素的扰动引起的。然而，如此大规模的改变不足以定义域形成的最低要求。此外，在多大程度上可以设计域才刚刚开始探索。为了创建接触域，我们将组织不变域边界下的 2 kb DNA 序列（包含 CTCF 结合位点 (CBS) 和转录起始位点 (TSS)）插入 11 个染色体的 16 个异位基因座中，并描述了它的建筑影响。根据局部限制，该片段可变地形成新域、划分现有域、改​​变区室化并启动反映染色质环挤出的接触。CBS 或 TSS 的删除单独或在插入内组合显示其对基因组折叠的独特贡献。总之，短 DNA 插入足以以受染色质背景影响的方式塑造空间基因组。