Rapid and Quantitative Functional Interrogation of Human Enhancer Variant Activity in Live Mice.

ABSTRACT: Functional analysis of non-coding variants associated with human congenital disorders remains challenging due to the lack of efficient in vivo models. Here we introduce dual-enSERT, a robust Cas9-based two-color fluorescent reporter system which enables rapid, quantitative comparison of enhancer allele activities in live mice of any genetic background. We use this new technology to examine and measure the gain- and loss-of-function effects of enhancer variants linked to limb polydactyly, autism, and craniofacial malformation. By combining dual-enSERT with single-cell transcriptomics, we characterize variant enhancer alleles at cellular resolution, thereby implicating candidate molecular pathways in pathogenic enhancer misregulation. We further show that independent, polydactyly-link

SUBMITTER: Hollingsworth EW 

PROVIDER: S-EPMC10723448 | biostudies-literature | 2023 Dec

REPOSITORIES: biostudies-literature