English (United Kingdom)

https://curated-unify.zendy.io/wp-json/zendy-region/v1/featured_content/oa?rat=en

https://curated-unify.zendy.io/wp-json/zendy-region/v1/highlighted_journal/

Global: Zendy Plus annual

Presents the access of premium content as premium feature

Premium Content

Presents the keyphrase highlighting as premium feature

Keyphrase Highlighting

Presents the summarisation as premium feature

Summarisation

Insights

Presents the pdf analysis as premium feature

PDF Analysis

Presents the zaia usage as premium feature

ZAIA

Global: Zendy Plus monthly

Global: Zendy Tools annual

Global: Zendy Tools monthly

Global: Zendy Free Plan

Background  Accumulation of somatic mutations caused by both endogenous and exogenous exposures is a high risk for human health, in particular, cancer. Efficient detection of somatic mutations is crucial for risk assessment of different types of exposures. Due to its requirement in the process of attaching glycosylphatidylinositol- (GPI-) anchored proteins to the cell surface, the  PIG-A  gene located on the X-chromosome is used in both in vivo and in vitro mutation assays. Loss-of-function mutations in  PIG-A  lead to the elimination of GPI-anchored proteins such that they can no longer be detected on the cell surface by antibodies. Historically, mutation assays based on the  PIG-A  gene rely on the staining of these cell-surface proteins by antibodies; however, as with any antibody-based assay, there are major limitations, especially in terms of variability and lack of specific antibodies.    Results  In the current study, we developed a modified  PIG-A  mutation assay that uses the expression of GPI-anchored fluorescent proteins (henceforth referred to as a GPI-sensor), whereby the presence of fluorescence on the cell membrane is dependent on the expression of  wild-type PIG-A . Using our modified  PIG-A  mutation assay, we have achieved complete separation of  wild type  cells and spontaneously mutated cells, in which the presence of  PIG-A  mutations has been confirmed via proaerolysin resistance and gene sequencing.    Conclusion  This study establishes a novel  PIG-A  mutation assay using GPI-anchored fluorescent protein expression that eliminates the need for antibody-based staining. This GPI-sensor  PIG-A  mutation assay should be widely applicable for accurate and efficient testing of genotoxicity for use in many mammalian and vertebrate cells.

Development of a novel PIG-A gene mutation assay based on a GPI-anchored fluorescent protein sensor