The DZIP3 Knockout HAP1 Polyclonal Cells product provides a CRISPR/Cas9-edited polyclonal knockout cell population with targeted disruption of the DZIP3 gene. This product offers a loss-of-function model to investigate DZIP3-dependent cellular processes. The polyclonal format ensures a diverse pool of edited alleles, enabling robust functional studies without clonal bias.
The knockout is engineered in the HAP1 cell line, a near-haploid fibroblast-like cell model derived from the KBM-7 chronic myeloid leukemia line. HAP1 cells are adherent and retain a haploid karyotype, making them ideal for genetic screening and gene-editing applications. Their haploid nature facilitates straightforward genotype-phenotype correlation and eliminates confounding effects from homologous chromosomes.
DZIP3 encodes an E3 ubiquitin ligase that critically regulates the Hedgehog signaling pathway by targeting GLI transcription factors (GLI1, GLI2, GLI3) for ubiquitin-mediated proteasomal degradation. This activity is inhibited by Hedgehog ligands such as Sonic hedgehog (SHH) through the canonical pathway involving PTCH1, SMO, and SUFU. Thus, DZIP3 functions as a negative regulator downstream of SUFU, controlling the stability of GLI proteins. Additionally, DZIP3 interacts with DAZ family proteins and ubiquitin-conjugating enzymes, playing roles in mRNA stability and spermatogenesis.
In the HAP1 background, DZIP3 knockout is particularly valuable for dissecting Hedgehog signaling dynamics. The haploid genetic context simplifies the analysis of pathway activation and GLI protein turnover. Because HAP1 cells are derived from a leukemia line, this model also enables cancer-related studies, including the role of aberrant Hedgehog signaling in oncogenesis. Researchers can use this system to evaluate how loss of DZIP3 influences cell proliferation, differentiation, and response to pathway agonists or antagonists.
This product is suitable for a variety of experimental approaches, including Gli-luciferase reporter assays to quantify Hedgehog pathway activity, Western blotting and immunofluorescence to monitor GLI protein levels, and ubiquitination assays to directly assess DZIP3 ligase function. Additionally, RT-qPCR of Hedgehog target genes, cell proliferation assays, and RNA-seq can provide comprehensive insights into the transcriptional and phenotypic consequences of DZIP3 loss. These applications support research in cancer biology, spermatogenesis, and drug target validation. For additional technical details, please contact Ascent Research.
