The HECTD3 Knockout HAP1 Polyclonal Cells consist of a CRISPR/Cas9-mediated polyclonal knockout population in which the HECTD3 gene has been disrupted in the near-haploid human HAP1 cell line. This pooled format preserves the diversity of editing events, enabling robust functional analysis without clonal isolation, and is ideally suited for assays that benefit from a representative loss-of-function model reflecting population-level responses.
HAP1 cells are a near-haploid adherent cell line originating from the KBM-7 chronic myeloid leukemia line, characterized by a single copy of most chromosomes. This haploid nature simplifies genetic perturbation and phenotypic interpretation, making HAP1 a well-established platform for high-throughput CRISPR screens, functional genomics, and drug target validation. The cells exhibit stable growth and efficient transfection, facilitating a wide range of molecular and cellular assays.
HECTD3 functions as an E3 ubiquitin ligase that catalyzes the ubiquitination and subsequent proteasomal degradation of key signaling proteins, notably MALT1 and Caspase-8. By controlling the stability of these substrates, HECTD3 acts as a negative regulator of NF-??B signaling downstream of TNF-?? and DNA damage-induced checkpoints, while also setting the threshold for apoptosis through Caspase-8 turnover. HECTD3 interacts directly with UBE2D family E2 enzymes to execute ubiquitin transfer and forms complexes with TRAF6, thereby integrating signals from p53-dependent pathways and modulating cell survival decisions. Representative pathway components influenced by HECTD3 include MALT1, TRAF6, NF-??B, Caspase-8, BAX, and p53.
Knockout of HECTD3 in the HAP1 background is expected to stabilize MALT1 and Caspase-8, leading to constitutive enhancement of NF-??B activity and a profound shift in apoptotic responsiveness. This cellular context is exceptionally suited for dissecting the crosstalk between ubiquitination-dependent proteolysis and programmed cell death, and for probing HECTD3??s contribution to oncogenic processes in cancers such as breast, gastric, and lung carcinomas, where dysregulated NF-??B and defective apoptosis are prevalent.
This polyclonal knockout product supports a diverse array of research applications, including functional genomics screens, mechanistic investigations into NF-??B and apoptosis signaling, ubiquitination studies, and cancer drug discovery efforts. Representative experimental approaches include Western blotting for substrate accumulation, RT-qPCR profiling of NF-??B target genes, Annexin V/PI apoptosis assays, NF-??B luciferase reporter assays, co-immunoprecipitation to map protein complexes, and chemotherapy sensitivity assays. For additional details, please contact Ascent Research.
