Scalable Dual-Fluorescence Assay for Functional Interpretation of HNF-4α Missense Variants.

ABSTRACT:

Aim

The study aimed to develop a scalable dual-fluorescence assay in cells to enable the functional interpretation of HNF-4α missense variants identified in exome sequencing, which can be used to guide clinical diagnosis.

Methods

Using mOrange2 and GFP fluorescence proteins to track the expression of HNF-4α (HNF-4α-mOrange2) and reporter activity under the control of the HNF-1α promoter (pHNF1A-GFP), respectively, we designed a dual-fluorescence assay to evaluate the expression level, cellular localization, and transcriptional function of HNF-4α simultaneously in live cells. To assess the scalable characteristic of the assay, a small library containing five previously reported mutations and wild-type HNF-4α was constructed. Cells infected with this library were sorted into d

SUBMITTER: Guo Y 

PROVIDER: S-EPMC8883583 | biostudies-literature | 2022

REPOSITORIES: biostudies-literature