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Updated: Jun 7, 2025

Using the E1A Minigene Tool to Study mRNA Splicing Changes
Published on: April 22, 2021
Minigene-based splice assays provide new insights on intronic variants of the PKHD1 gene
Yiyin Zhang1, Ran Zhang1, Xiaomeng Shi1
1Department of Nephrology, The Affiliated Municipal Hospital of Qingdao University, No.5 Donghai Middle Road, Qingdao, 266071, China.
Background:
Autosomal Recessive Polycystic Kidney Disease (ARPKD) is a rare hereditary disorder caused by variants in PKHD1. Currently, aberrant splicing has been reported to play important roles in genetic disease. Our goal is to analyze intronic variants in PKHD1 at the mRNA level.
Results:
The 12 candidate variants were introduced into the corresponding minigene and functionally assayed in HEK 293T and Hela cells. We identified 11 variants that induce splicing alterations, resulting in various consequences such as skipping of exons, intron retention and protein truncation.
Conclusions:
This underlined the importance of mRNA-level assessment for genetic diagnostics in related genetic disorders.
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