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Rapid and Functional Quantitative Interrogation of Human Enhancer Variants in Live Mice

ABSTRACT: Functional characterisation of noncoding variants linked to human congenital disorders remains challenging due to the lack of efficient in vivo models. Here we introduce dual-enSERT, a robust two-color fluorescent reporter system which enables rapid comparative assessment of human enhancer variant activities in live mice. We use this new technology to examine the gain- and loss-of-function effects of enhancer variants previously linked to limb polydactyly, autism, and craniofacial malformation. By combining dual-enSERT with single-cell transcriptomics, we characterize variant enhancer allele activity at cellular resolution, revealing candidate molecular pathways implicated in pathogenic enhancer misregulation. We also use dual-enSERT to show that independent polydactyly-linked enhancer variants lead to ectopic expression in the same cell populations, indicating shared genetic mechanisms underlying noncoding variant pathogenesis. Finally, we streamline dual-enSERT for F0 analysis by placing both reporters on the same allele separated by an insulator. Our F0 dual-enSERT cuts the experimental time of noncoding variant selection to corresponding comparative in vivo activity in live embryos from three months to two weeks.

ORGANISM(S): Mus musculus

PROVIDER: GSE244244 | GEO | 2024/01/18

REPOSITORIES: GEO

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