The CCNA1 Knockout HAP1 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population designed to disrupt the CCNA1 gene in HAP1 cells. This product provides a heterogeneous pool of edited cells, enabling loss-of-function studies of Cyclin A1 without clone selection. The polyclonal format ensures robust, reproducible readouts across a population of knockout cells, suitable for diverse experimental workflows.
HAP1 is a human near-haploid chronic myeloid leukemia (CML) cell line, derived from the myeloid lineage and originating from KBM-7 cells. With a fibroblast-like morphology and a single copy of most chromosomes, HAP1 offers a genetically simplified system where gene disruption directly yields a null phenotype. This feature makes it ideal for knockout screens and rapid genotype?Cphenotype analysis in drug discovery and functional genomics.
Cyclin A1, encoded by CCNA1, serves as a regulatory subunit for CDK2 and CDK1, driving G1/S and G2/M transitions via phosphorylation of RB1, which releases E2F1 to activate S-phase genes like CDC6 and MCM2-7. CCNA1 expression is activated by MYB, E2F1, and repressed by p53, while its activity is inhibited by CDKN1A (p21) and CDKN1B (p27) and modulated by INCA1 and SKP2. Knockout of CCNA1 ablates Cyclin A1/CDK function, preventing RB1 phosphorylation and E2F-mediated transcription, ultimately causing G1 arrest.
In HAP1 cells, haploidy ensures complete loss of Cyclin A1 function, making this polyclonal knockout a potent model for studying cell cycle dysregulation in leukemia. CCNA1 is implicated in acute myeloid leukemia and testicular cancer, and its disruption in this background allows exploration of Cyclin A1-dependent proliferation pathways and p53 interplay. The model facilitates investigation of hematopoietic stem cell regulation and meiosis mechanisms.
Researchers can employ flow cytometry with PI or BrdU staining to assess cell cycle arrest, western blotting for Cyclin A1, CDK2, and phospho-RB, and proliferation assays (MTS/CCK-8) to evaluate growth inhibition. RT-qPCR of E2F target genes confirms downstream effects. Applications include drug target validation in leukemia, functional genomics screens, and meiosis research. For further details, contact Ascent Research.
