美国劳伦斯伯克利国家实验室Diane E. Dickel、Axel Visel等研究人员合作利用全面体内筛查揭示了人类增强子变异的表型影响。相关论文于2020年3月12日在线发表在《细胞》杂志上。
Title: Comprehensive In Vivo Interrogation Reveals Phenotypic Impact of Human Enhancer Variants
Author: Evgeny Z. Kvon, Yiwen Zhu, Guy Kelman, Catherine S. Novak, Ingrid Plajzer-Frick, Momoe Kato, Tyler H. Garvin, Quan Pham, Anne N. Harrington, Riana D. Hunter, Janeth Godoy, Eman M. Meky, Jennifer A. Akiyama, Veena Afzal, Stella Tran, Fabienne Escande, Brigitte Gilbert-Dussardier, Nolwenn Jean-Marais, Sanjarbek Hudaiberdiev, Ivan Ovcharenko, Matthew B. Dobbs, Christina A. Gurnett, Sylvie Manouvrier-Hanu, Florence Petit, Axel Visel, Diane E. Dickel, Len A. Pennacchio
Abstract: Establishing causal links between non-coding variants and human phenotypes is an increasingchallenge. Here, we introduce a high-throughput mouse reporter assay for assessingthe pathogenic potential of human enhancer variants in vivo and examine nearly a thousand variants in an enhancer repeatedly linked to polydactyly.We show that 71% of all rare non-coding variants previously proposed as causal leadto reporter gene expression in a pattern consistent with their pathogenic role. Variantsobserved to alter enhancer activity were further confirmed to cause polydactyly inknockin mice. We also used combinatorial and single-nucleotide mutagenesis to evaluatethe in vivo impact of mutations affecting all positions of the enhancer and identified additionalfunctional substitutions, including potentially pathogenic variants hitherto not observedin humans. Our results uncover the functional consequences of hundreds of mutationsin a phenotype-associated enhancer and establish a widely applicable strategy forsystematic in vivo evaluation of human enhancer variants.