The ING2 Knockout HAP1 Polyclonal Cells represent a CRISPR/Cas9-edited polyclonal knockout cell population derived from the HAP1 human cell line, specifically designed for targeted disruption of the ING2 gene. This polyclonal format captures a spectrum of editing events, enabling functional analysis without the biases of clonal isolation. The knockout model serves as a versatile tool for dissecting ING2-dependent pathways in a near-haploid background.
HAP1 cells are a near-haploid human cell line originating from the KBM-7 chronic myeloid leukemia line, characterized by a single copy of most chromosomes. This genetic configuration ensures that CRISPR/Cas9-mediated gene disruption yields a uniform loss-of-function phenotype, making HAP1 an ideal host for genetic perturbation screens and tumor suppressor studies. The cell line retains signaling networks relevant to leukemia biology, providing a physiologically appropriate system for cancer research.
ING2 is a tumor suppressor that functions as a scaffold linking histone modifications to TP53 activation. Via its PHD finger, ING2 binds H3K4me3 at chromatin and recruits histone acetyltransferases such as PCAF and p300, leading to acetylation and stabilization of TP53. Activated TP53 then drives transcription of CDKN1A (p21) and BAX, promoting cell cycle arrest and apoptosis. ING2 exists in multi-protein complexes with ING1, ING3, SAP30, Sin3A, HDAC1, HDAC2, and RBP1, and integrates signals from upstream regulators including TP53 itself, E2F transcription factors, TGF-beta, and DNA damage. These interactions position ING2 as a key coordinator of TP53 signaling, histone acetylation, and tumor suppression.
Disruption of ING2 in the near-haploid HAP1 context unmasks its tumor-suppressive activities, enabling precise dissection of how ING2 loss affects TP53 acetylation, downstream target expression, and apoptotic responses. The monogenic knockout enhances the clarity of phenotypic outcomes, making it particularly valuable for studying the intersection of chromatin regulation and apoptosis in a leukemia-derived model.
These knockout cells are suitable for a variety of applications, including western blotting for TP53, p21, and cleaved caspase-3; RT-qPCR of TP53 target genes; co-immunoprecipitation of ING2 complexes; ChIP-qPCR to assess histone marks; flow cytometry for cell cycle and apoptosis; and viability assays for drug sensitivity profiling. Typical research areas include tumor suppressor biology, apoptosis signaling, histone modification mechanisms, and CRISPR-based functional genomics. For further information, please contact Ascent Research.
