DZIP3 Knockout HEK293T Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population derived from the HEK293T human embryonic kidney cell line. These polyclonal cells harbor targeted disruption of the DZIP3 gene, encoding an E3 ubiquitin-protein ligase with a RING finger domain. The CRISPR/Cas9-mediated gene disruption generates a heterogeneous pool of knockout variants, enabling loss-of-function studies without single-cell cloning. This model provides a physiologically relevant system to investigate DZIP3-dependent regulation of RNA metabolism and protein degradation within the context of a widely used, highly transfectable mammalian cell host.
HEK293T cells are human embryonic kidney cells immortalized with sheared adenovirus type 5 DNA. They grow adherently and constitutively express the SV40 large T antigen, which drives efficient episomal replication of plasmids containing the SV40 origin of replication. The cell line is renowned for its high transfection efficiency, making it a preferred system for transient protein expression, viral vector production, and functional genomic screens. The stable, well-characterized nature of HEK293T cells provides a robust backdrop for dissecting the molecular functions of E3 ubiquitin ligases such as DZIP3.
DZIP3 functions as an E3 ubiquitin ligase within the ubiquitin-proteasome system, coupling ubiquitin to substrate proteins to signal their degradation or alter their activity. Mechanistic studies indicate that DZIP3 interacts with the DAZ and DAZL RNA-binding proteins, and likely with other RNA-binding proteins, to regulate RNA metabolic processes. DZIP3 may ubiquitinate components of ribonucleoprotein complexes, thereby controlling mRNA stability, splicing, or transport. Through these interactions, DZIP3 is thought to coordinate post-transcriptional gene regulation essential for germ cell development and spermatogenesis. The pathway involves key components such as ubiquitin, the proteasome, and RNA-binding proteins, positioning DZIP3 at the intersection of proteostasis and RNA biology.
In HEK293T cells, the DZIP3 knockout polyclonal pool allows researchers to examine how loss of this ligase affects cellular ubiquitination profiles, RNA metabolism, and protein turnover in a non-germline, easily manipulable system. Although HEK293T cells are of kidney origin, they express many core components of mRNA regulation and the ubiquitin-proteasome machinery, enabling investigation of broadly conserved mechanisms. The polyclonal nature of the knockout population models heterogeneous gene disruption, which can unmask dominant-negative effects or incomplete penetrance of phenotypes, providing a more nuanced view of DZIP3 function than clonal cell lines.
Typical applications include studying ubiquitin-mediated regulation of RNA processing, investigating male infertility mechanisms, and functional genomics of E3 ligases. Representative assays with this model include Sanger sequencing to verify genomic edits, western blotting to assess DZIP3 protein levels, RT-qPCR to monitor mRNA expression, co-immunoprecipitation to probe interactions with DAZ and DAZL, and ubiquitination assays to track substrate modification. Immunofluorescence can reveal changes in protein localization. This product supports rigorous dissection of DZIP3??s roles in both normal physiology and disease states such as male infertility and cancer. For further details or technical support, please contact Ascent Research.
