The HMG20A Knockout HAP1 Polyclonal Cells consist of a CRISPR/Cas9-edited polyclonal pool in which the HMG20A gene has been disrupted, generating a loss-of-function model for investigating HMG20A-dependent transcriptional regulation. This population-based format incorporates multiple independent editing events, reducing clonal artefacts and providing a robust system for population-level assays. The knockout cells are particularly suited for studying the role of HMG20A in neuronal gene silencing and cell cycle modulation within a human cellular context.
HAP1 is a human near-haploid cell line derived from KBM-7 chronic myeloid leukemia cells, characterized by an adherent, fibroblast-like morphology. Its haploid karyotype simplifies CRISPR-based gene disruption, as single-copy knockout events yield unambiguous phenotypic consequences. HAP1 cells retain functional disease-relevant pathways and are widely used in functional genomics, drug response profiling, and high-throughput screens, providing a standardized background for gene-edited models.
HMG20A (BRAF35) is a core subunit of the BRAF35-HDAC corepressor complex that mediates transcriptional repression of neuronal genes in non-neuronal cells. It bridges the master repressor REST with CoREST (RCOR1), histone deacetylase HDAC1, and the lysine-specific demethylase LSD1 (KDM1A), alongside PHF21A. This assembly coordinates deacetylation and demethylation of histone H3 at promoters of REST target genes, such as SYN1 and SCN2A, maintaining a transcriptionally silent chromatin state. HMG20A is regulated by REST and neuronal transcription factors; its disruption derepresses neuronal gene programs and can influence cell cycle progression.
Within the HAP1 near-haploid background, HMG20A knockout creates a powerful tool to dissect REST-mediated silencing. Minimal genetic redundancy amplifies phenotypic consequences, facilitating analysis of neuronal gene de-repression. This model is especially valuable for neurodevelopmental and autism spectrum disorder research, where disrupted chromatin remodeling is implicated. Monitoring cell cycle dynamics and neuron-specific gene activation helps probe links between epigenetic regulation and disease.
Applications include Western blotting, RT?qPCR, RNA?seq, and ChIP?qPCR to confirm knockout and assess transcriptomic and epigenomic changes. Immunofluorescence and reporter assays enable pathway visualization. The polyclonal cells are suitable for high-throughput screening targeting the REST?CoREST?HMG20A?HDAC/LSD1 axis, relevant to neurological drug discovery. For further information, contact Ascent Research.
