The ALKBH2 Knockout HAP1 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population derived from the HAP1 near-haploid human cell line. This product features a targeted gene disruption of ALKBH2, creating a loss-of-function model for investigating DNA repair and the maintenance of genomic integrity. As a polyclonal population, it offers a cost-effective and rapidly deployable tool for functional studies without requiring single-cell clonal isolation. ALKBH2 encodes a DNA repair dioxygenase that directly reverses alkylation damage, and its knockout enables detailed dissection of the base excision repair pathway and cellular responses to genotoxic stress.
The host cell line, HAP1, originates from a near-haploid chronic myeloid leukemia (CML) cell line derived from the male KBM-7 line. HAP1 cells harbor the BCR-ABL oncogenic fusion, providing a leukemia-relevant context for cancer biology and drug sensitivity research. The near-haploid karyotype, with only one copy of most chromosomes, greatly facilitates genetic perturbation and phenotypic analysis by eliminating confounding heterozygosity. This unique genomic architecture makes HAP1 an ideal platform for knockout-based screening and mechanistic dissection of gene function, ensuring high-confidence genotype-phenotype correlations.
ALKBH2 is an ??-ketoglutarate-dependent dioxygenase that catalyzes the oxidative demethylation of 1-methyladenine and 3-methylcytosine in DNA, directly reversing methylation damage. Expression of ALKBH2 is regulated upstream by the tumor suppressor TP53 in response to DNA damage. ALKBH2 interacts with proliferating cell nuclear antigen (PCNA) to be recruited to replication foci during S-phase, coupling repair to DNA synthesis. This interaction ensures timely removal of alkylation lesions, promoting restored DNA bases, reduced mutagenesis, and overall genomic stability. The pathway involves base excision repair enzymes and alkylated DNA substrates, with ALKBH2 and PCNA as central components coordinating damage recognition and resolution.
In the HAP1 cellular background, disruption of ALKBH2 creates a model highly susceptible to alkylation-induced DNA damage, enabling rigorous study of repair deficiencies and synthetic lethal interactions. The presence of BCR-ABL adds clinical relevance, as it reflects Philadelphia chromosome-positive leukemias with potentially altered DNA damage responses. This knockout model is well-suited for examining how ALKBH2 deficiency impacts cell cycle progression, apoptosis, and chromosomal integrity in a malignant context. Moreover, the haploid genome allows unambiguous assessment of DNA repair kinetics without compensatory allele effects.
This polyclonal knockout cell population supports diverse applications in DNA damage research. Researchers can employ Western blotting, RT-qPCR, and immunofluorescence to confirm ALKBH2 disruption. Functional assays such as the comet assay, ??-H2AX foci analysis, and cell viability tests with alkylating agents (e.g., methyl methanesulfonate) quantify DNA damage and repair efficiency. The model is suitable for CRISPR-based synthetic lethality screens to identify genes enhancing alkylation sensitivity, and for developing DNA repair inhibitors. For further details, please contact Ascent Research.
