CRISPR screen links BAF mutation to premature ageing

A genome-wide CRISPR screen in fibroblasts from patients with Néstor-Guillermo Progeria Syndrome (NGPS) identified 43 genes that mitigate multiple cellular defects associated with the disease. The findings suggest that the A12T mutation in BAF disrupts protein synthesis, contributing to cellular dysfunction and premature ageing.

By: Gorm Palmgren - Feb. 17, 2025

NGPS is a rare premature ageing disorder caused by a mutation in BANF1, encoding barrier-to-autointegration factor (BAF), a nuclear envelope-associated protein. Using patient-derived fibroblasts, researchers conducted a CRISPR-Cas9-based synthetic rescue screen to identify genes whose loss could reverse key NGPS cellular phenotypes.

Figure 1. A Schematic detailing the four main steps involved in the whole genome CRISPR screening... — Figure 1. A Schematic detailing the four main steps involved in the whole genome CRISPR screening set-up in NGPS broblasts. Reproduced under the Creative Commons license from Figure 2 in Breusegem et al. (2024) Nature Communications, doi.org/10.1038/s41467-025-56916-5

The CRISPR screen was designed to assess the effect of gene knockouts on four NGPS-specific phenotypes: nuclear shape abnormalities, mislocalisation of emerin, increased nuclear envelope ruptures, and micronuclei formation (see Figure 1). The researchers employed a microscopy-based approach to quantify these features in fibroblasts stably expressing Cas9.

A whole-genome arrayed CRISPR library targeting ~20,000 human genes was used, with hits selected based on their ability to normalise multiple phenotypes. Validation experiments using siRNA knockdowns and protein synthesis assays confirmed the functional relevance of the identified genes. Further analysis linked several hits to human ageing traits through genome-wide association studies, reinforcing their potential role in ageing-related cellular deterioration.

The identified genes were enriched in pathways related to protein synthesis, RNA transport, and osteoclast development, consistent with NGPS symptoms such as skeletal abnormalities and lipoatrophy. Further validation in a Caenorhabditis elegans NGPS model demonstrated that knockdown of selected genes improved viability, supporting their relevance to disease pathology.

Notably, the study found that NGPS cells exhibited an increased rate of protein synthesis and translation errors, which may accelerate ageing-associated cellular decline. Inhibiting protein synthesis pharmacologically or via gene knockdown improved nuclear envelope integrity, highlighting potential therapeutic targets.

This research was led by Sophia Y. Breusegem and Delphine Larrieu at the University of Cambridge and colleagues at several European institutions. The study was published in Nature Communications on 16 February 2025.

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