NanoLuc reporters identify COL4A5 nonsense mutations susceptible to drug-induced stop codon readthrough
Abstract via Europe PMC
Alport syndrome, a disease of kidney, ear, and eye, is caused by pathogenic variants in the COL4A3 , COL4A4 , or COL4A5 genes encoding collagen α3α4α5(IV) of basement membranes. Collagen IV chains that are truncated due to nonsense variants/premature termination codons (PTCs) cannot assemble into heterotrimers or incorporate into basement membranes. To investigate the feasibility of PTC readthrough therapy for Alport syndrome, we utilized two NanoLuc reporters in transfected cells: full-length for monitoring translation, and a split version for assessing readthrough product function. Full-length assays of 49 COL4A5 nonsense variants identified eleven as susceptible to PTC readthrough using various readthrough drugs. In split-NanoLuc assays, the predicted missense α5(IV) readthrough products of five nonsense mutations could heterotrimerize with α3(IV) and α4(IV). Readthrough was also observed in kidney cells from an engineered Col4a5 PTC mouse model. These results suggest that readthrough therapy is a feasible approach for a fraction of patients with Alport syndrome.
Evidence snapshot note
This reference is cited by the validated MVA · BUB1B evidence snapshot (data as of 2026-10-17) as Omachi et al. — COL4A5 readthrough reporters.
- Model in the source
- HEK293 COL4A5 cDNA reporters; different gene and context
- Where in the publication
- Figure 6: gentamicin plus mefloquine
- Cited by evidence records
- MVA-RT-002
The note describes how the snapshot used the source; it is not a summary of the publication's conclusions.
Identifiers and provenance
- DOI
10.1016/j.isci.2022.103891- PMID
35243249- PMCID
PMC8866893- Metadata from
- Crossref, Europe PMC, OpenAlex, fetched
- License (as declared by the source)
- https://www.elsevier.com/tdm/userlicense/1.0/
- Open access
- Yes
- Added
- from the MVA evidence snapshot
In the library
- Collections
- MVA Hackathon 2026 — BUB1B
- Tags
- None yet
- Keywords from the source
- Molecular biology, Biochemistry, Molecular Medicine