The impact of genomic distance on enhancer-promoter interactions at the CFTR locus.

Autor: Kerschner JL; Department of Genetics and Genome Sciences, Case Western Reserve University, Cleveland, Ohio, USA., Meckler F; Department of Genetics and Genome Sciences, Case Western Reserve University, Cleveland, Ohio, USA., Coatti GC; Department of Genetics and Genome Sciences, Case Western Reserve University, Cleveland, Ohio, USA., Vaghela N; Department of Genetics and Genome Sciences, Case Western Reserve University, Cleveland, Ohio, USA., Paranjapye A; Department of Genetics and Genome Sciences, Case Western Reserve University, Cleveland, Ohio, USA., Harris A; Department of Genetics and Genome Sciences, Case Western Reserve University, Cleveland, Ohio, USA.
Jazyk: angličtina
Zdroj: Journal of cellular and molecular medicine [J Cell Mol Med] 2024 Feb; Vol. 28 (4), pp. e18142.
DOI: 10.1111/jcmm.18142
Abstrakt: We identified and characterized multiple cell-type selective enhancers of the CFTR gene promoter in previous work and demonstrated active looping of these elements to the promoter. Here we address the impact of genomic spacing on these enhancer:promoter interactions and on CFTR gene expression. Using CRISPR/Cas9, we generated clonal cell lines with deletions between the -35 kb airway enhancer and the CFTR promoter in the 16HBE14o - airway cell line, or between the intron 1 (185 + 10 kb) intestinal enhancer and the promoter in the Caco2 intestinal cell line. The effect of these deletions on CFTR transcript abundance, as well as the 3D looping structure of the locus was investigated in triplicate clones of each modification. Our results indicate that both small and larger deletions upstream of the promoter can perturb CFTR expression and -35 kb enhancer:promoter interactions in the airway cells, though the larger deletions are more impactful. In contrast, the small intronic deletions have no effect on CFTR expression and intron 1 enhancer:promoter interactions in the intestinal cells, whereas larger deletions do. Clonal variation following a specific CFTR modification is a confounding factor particularly in 16HBE14o - cells.
(© 2024 The Authors. Journal of Cellular and Molecular Medicine published by Foundation for Cellular and Molecular Medicine and John Wiley & Sons Ltd.)
Databáze: MEDLINE
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