HEATR3 Knockout HAP1 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population in which the human HEATR3 gene has been disrupted using a CRISPR/Cas9-mediated gene-editing approach. This product provides a genetically heterogeneous pool of HAP1 cells bearing loss-of-function alleles of HEATR3, enabling pooled functional genomics studies and high-throughput screening applications. The polyclonal format captures a range of editing outcomes, offering a representative model for studying the gene’s role without the clonal artifacts that can arise from single-cell isolation.
The host cell line, HAP1, is a near-haploid human cell line derived from the chronic myeloid leukemia line KBM-7. It retains a haploid chromosome set except for disomic regions on chromosomes 8 and 15. This near-haploid karyotype simplifies gene knockout studies because disruption of a single allele can result in a functional null phenotype. HAP1 cells are widely employed for CRISPR-based functional screens, genetic interaction mapping, and mechanistic studies due to their robust growth, ease of editing, and well-characterized genetic background.
HEATR3 encodes a nucleolar protein that functions in the late stages of 60S ribosomal subunit maturation. It localizes to the nucleolus and participates in pre-60S particle assembly, facilitating processing of 28S rRNA and enabling efficient protein synthesis. HEATR3 is regulated upstream by MYC and mTORC1 signaling, which govern ribosome biogenesis in response to growth and nutrient cues. It interacts with critical ribosomal biogenesis factors including NPM1, NCL, RPL5, RPL11, NOP2, and PES1. Loss of HEATR3 disrupts pre-rRNA processing intermediates, impairs 60S subunit formation, and reduces global translation, ultimately suppressing cell growth and proliferation.
In the HAP1 context, HEATR3 knockout provides a powerful system to dissect ribosome biogenesis pathways with minimal genetic redundancy. The near-haploid background ensures that gene disruption yields a clear loss-of-function phenotype, facilitating dose-response studies and synthetic lethality screens. This model is particularly relevant for cancer research, as MYC-driven ribosomal gene expression is a hallmark of many malignancies, and HEATR3 may represent a vulnerability in cells with high translational demand. Additionally, the model allows investigation of nucleolar stress responses and cross-talk between ribosome assembly and cell cycle checkpoints.
This polyclonal knockout cell population is suitable for a wide range of research applications, including functional dissection of ribosome biogenesis, cancer dependency mapping, identification of synthetic lethal interactions, and drug target validation for ribosome-targeted therapies. Representative experimental approaches include Western blotting of ribosomal proteins, RT-qPCR for pre-rRNA species, polysome profiling to assess translation efficiency, puromycin incorporation assays, nucleolar stress immunofluorescence, and proliferation and viability measurements. For further information or to discuss custom requirements, please contact Ascent Research.
