Disrupting the three-dimensional regulatory topology of the Pitx1 locus results in overtly normal development

• DOI: 10.1242/dev.158550
• Publication Date: 2018-03-16T11:35:26Z
• Authors: Richard Sarro; Acadia A. Kocher; Deena Emera; Severin Uebbing; Emily V. Dutrow; Scott D. Weatherbee; Timothy Nottoli; James P. Noonan
• Keywords: Chromosome conformation capture

ABSTRACT Developmental gene expression patterns are orchestrated by thousands of distant-acting transcriptional enhancers. However, identifying enhancers essential for the their target genes has proven challenging. Maps long-range regulatory interactions may provide means to identify crucial developmental expression. To investigate this hypothesis, we used circular chromosome conformation capture coupled with interaction maps in mouse limb characterize topology Pitx1, which is hindlimb development. We identified a robust hindlimb-specific between Pitx1 and putative enhancer. interrogate role regulation, genome editing delete enhancer mouse. Although deletion completely disrupts interaction, only mildly affected, without any detectable compensatory promoter potentially redundant null mice did not exhibit characteristic morphological defects Pitx1−/− mutant. Our results suggest that robust, tissue-specific physical at have limited predictive value mutations strong loss-of-function phenotypes.