The CACUL1 Knockout HAP1 Polyclonal Cells provide a CRISPR/Cas9-edited polyclonal knockout cell population with targeted disruption of the CACUL1 gene. This heterogeneous pool of cells offers a robust loss-of-function model for studying CACUL1-dependent processes, avoiding clonal artifacts. The polyclonal format is well-suited for bulk assays and functional genomics applications.
The HAP1 cell line is a near-haploid human hematopoietic model derived from KBM-7 chronic myeloid leukemia cells. Its haploid karyotype permits efficient gene disruption and simplifies phenotypic analysis, as only a single allele needs targeting. HAP1 cells retain critical signaling networks of myeloid origin, making them relevant for cancer and cell cycle research.
CACUL1 (CDK2-associated cullin domain 1) encodes a cullin-related protein that binds CDK2 and is thought to inhibit its kinase activity, thereby controlling G1/S progression. The gene is transcriptionally regulated by E2F1, integrating it into a regulatory network involving Cyclin E, CDK2, RB1, and E2F1. CACUL1’s cullin domain suggests potential involvement in ubiquitin-mediated processes, though its best-characterized function is negative regulation of CDK2. Disruption of CACUL1 releases CDK2 inhibition, enhancing RB1 phosphorylation and promoting E2F-dependent transcription, which accelerates cell cycle entry.
In the context of HAP1 cells, which originate from a CML background, CACUL1 knockout provides a tool for dissecting cell cycle deregulation in hematopoietic malignancies. The near-haploid nature ensures unambiguous phenotypes, facilitating dose-response studies and synthetic lethality screens. This model enables investigation of how CACUL1 loss cooperates with oncogenic drivers or influences sensitivity to CDK inhibitors.
Applications include Western blot analysis of CDK2 substrates such as phospho-RB1, flow cytometric cell cycle profiling, and RT-qPCR quantification of E2F target genes. Immunofluorescence detection of phospho-RB1 enables cell-level resolution of cell cycle status. These cells are also suitable for functional genomics screens and drug sensitivity assays. For further information, please contact Ascent Research.
