Assay for identification of 5′ UTR mutations impacting translation and protein production from NDD genes

5′ untranslated regions (UTRs) regulate the translation of the coding sequences and are often mutated in patients with autism and neurodevelopmental disorders (NDDs). However, their functional impacts are unclear, especially in the brain. 

Here, the researchers benchmarked approaches for functionally characterizing hundreds of UTR variants, including high-throughput assays in the living brain.

The authors  identified >100 altering translation, with a subset subsequently altering endogenous protein production in patient lymphoblastoid cell lines using massively parallel reporter assay (MPRA).

They optimized Cre-dependent MPRAs for assessment in neurons in vivo and demonstrate that neurons have different principles of regulation by 5′ UTRs and discover mutations altering translational activity. 

They found protein translation changes only correlated with altering UTR structure.

https://www.cell.com/cell-reports-methods/fulltext/S2667-2375(25)00283-8

https://sciencemission.com/UTR-mutations-impacting-translation