The ITCH Knockout HAP1 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population derived from the human HAP1 cell line, designed to disrupt the ITCH gene. This gene-edited product provides a loss-of-function model for investigating the E3 ubiquitin ligase ITCH, enabling detailed studies of its roles in ubiquitin-mediated proteolysis and cellular signaling without the confounding effects of a second allele.
The HAP1 cell line is a near-haploid human male cell model originating from a chronic myeloid leukemia (CML) patient. Its haploid karyotype simplifies genetic analyses, making it a robust platform for knockout studies, drug screening, and functional genomics. HAP1 cells retain core signaling pathways, including NF-??B and JNK cascades, and are widely adopted for investigating immune regulation, apoptosis, and cancer-related processes.
ITCH functions as a HECT-type E3 ubiquitin ligase that catalyzes ubiquitination and proteasomal degradation of substrate proteins, including c-FLIP, p63, p73, Notch1, Smad2, Smad3, JunB, and TXNIP. ITCH activity is regulated by upstream kinases JNK and MEKK1, as well as by inflammatory stimuli such as TNF-?? and IL-1. It interacts with adaptor proteins N4BP1, 14-3-3, and ??-arrestin, and collaborates with the E2 ubiquitin-conjugating enzyme UbcH7. Through these interactions, ITCH acts as a negative regulator of NF-??B and JNK signaling, modulating immune responses, apoptosis, and cell proliferation.
Disruption of ITCH in the haploid HAP1 background eliminates the complexity of diploid genetics, enabling unambiguous genotype-phenotype correlations. This model is particularly valuable for dissecting ubiquitin-proteasome system dynamics and their impact on immune signaling and cell growth control. Loss of ITCH function in HAP1 cells can reveal alterations in NF-??B and JNK signaling activity, as well as changes in the stability of key substrates, providing a clean genetic system for mechanistic inquiries.
Researchers can employ this ITCH knockout population to assess the ubiquitination and degradation of substrates such as c-FLIP and p63 using western blotting and ubiquitination assays. NF-??B reporter assays and JNK phosphorylation analyses provide functional readouts of signaling activities, while apoptosis assays and flow cytometry enable characterization of cell death responses. This model also supports drug screening campaigns aimed at identifying modulators of ITCH activity or its downstream pathways in autoimmune disease and cancer contexts. For further information or to request a quotation, please contact Ascent Research.
