The IKBKB Knockout HAP1 Polyclonal Cells constitute a genetically disrupted cell population generated by CRISPR/Cas9-mediated targeting of the IKBKB gene in the HAP1 cell line. This product provides a polyclonal knockout model of IKK??, the catalytic subunit of the I??B kinase complex, enabling functional studies of NF-??B signaling without clonal variation artifacts. The polyclonal population preserves a heterogeneous gene-edited background, reflecting diverse loss-of-function mutations across the cell pool. Researchers can employ this model to interrogate IKK??-dependent pathways in a haploid genetic context.
The HAP1 cell line is a near-haploid human cell line derived from a male patient with chronic myeloid leukemia in blast crisis. Its haploid karyotype simplifies genetic manipulation and phenotype interpretation, as single gene disruptions often result in unambiguous loss-of-function phenotypes. HAP1 cells retain many signaling networks relevant to hematological malignancies and are widely adopted in functional genomics screens, drug sensitivity assays, and pathway dissection. The haploid nature eliminates confounding effects from heterozygous mutations, making it an ideal host for knockout studies requiring clear genotype-phenotype correlations.
IKBKB encodes IKK??, a serine/threonine kinase that, together with IKK?? (CHUK) and the regulatory subunit NEMO (IKBKG), forms the IKK complex. Upon stimulation by upstream regulators such as TNF??, IL-1??, lipopolysaccharide (LPS), or TCR/BCR engagement, adaptors including TRADD, TRAF2, RIP1, and TAK1?CTAB2/3 transduce signals to the IKK complex. Activated IKK?? phosphorylates I??B??, triggering its ubiquitination and proteasomal degradation. This releases NF-??B dimers (typically p50/p65) to translocate to the nucleus and drive transcription of target genes involved in inflammation, cell survival, and proliferation, including IL-6, IL-8, TNF??, Bcl-2, Bcl-xL, and cyclin D1. Disruption of IKBKB thus abolishes signal-induced NF-??B activation.
In the HAP1 leukemic background, knockout of IKBKB uncouples IKK?? from downstream NF-??B responses, providing a powerful system to dissect IKK?¡?s role in hematopoietic cell survival and transformation. The haploid genome ensures that the functional consequences of IKBKB disruption are directly attributable to loss of IKK?? activity, without compensation from a second allele. This model is particularly relevant for studying the reliance of leukemic cells on NF-??B-driven prosurvival programs and for testing targeted therapies that inhibit IKK??. Additionally, it offers a clean background to explore cross-talk between NF-??B and other oncogenic pathways operative in CML blast crisis.
NF-??B pathway activation can be assessed by Western blot of phospho-I??B?? and phospho-p65, while target gene induction is measured by RT-qPCR of IL-6, IL-8, or TNF??. NF-??B luciferase reporter assays quantify transcriptional activity, and immunocytochemistry visualizes p65 nuclear translocation. Cytokine secretion is profiled by ELISA. Functional studies include flow cytometric apoptosis assays and cell viability measurements following IKK?? inhibitor treatment. Co-immunoprecipitation can probe IKK complex integrity. These cells support inflammatory disease modeling, drug sensitivity screens, and mechanistic studies of IKK?? in cancer and immune signaling. For further details, please contact Ascent Research.
