WDR33 Knockout HEK293T Cell Line

The WDR33 Knockout HEK293T Cell Line is a CRISPR/Cas9-edited knockout cell line targeting the WDR33 gene. This loss-of-function model enables dissection of WDR33’s role in pre-mRNA 3′ end processing by the cleavage and polyadenylation specificity factor (CPSF) complex, facilitating study of transcription termination and mRNA maturation in a human cellular system. By eliminating functional WDR33, it provides a defined background for mechanistic investigations.

HEK293T cells are a widely used human embryonic kidney epithelial line derived from HEK293 cells by stable expression of SV40 large T-antigen, enabling high episomal replication and robust protein expression. Characterized by rapid growth, efficient transfection, and a well-characterized transcriptome, HEK293T is ideal for gene editing studies. Its use as the parental line for WDR33 knockout ensures compatibility with standard workflows and direct comparison to wild-type controls in 3′ processing assays.

WDR33 encodes a core subunit of the cleavage and polyadenylation specificity factor (CPSF) complex that directly binds the AAUAAA polyadenylation signal. It nucleates CPSF assembly together with CPSF1 (CPSF160), CPSF4 (CPSF30), Fip1, and other subunits, and cooperates with cleavage stimulation factor (CstF), cleavage factors I/II (CFI/II), symplekin, and poly(A) polymerase (PAP) to catalyze endonucleolytic cleavage and polyadenylation. Disruption of WDR33 impairs CPSF complex integrity and recruitment of downstream factors, leading to defective 3′ end formation and altered global polyadenylation patterns. Such dysregulation is associated with cancer and neurodevelopmental disorders.

In the HEK293T background, WDR33 knockout provides a well-controlled system to probe the functional architecture of the mRNA 3′ processing machinery. The parental line??s rapid proliferation and high transfection efficiency enable acute readouts of polyadenylation defects via reporter constructs or genome-wide sequencing. Because HEK293T cells express a range of CPSF components, the knockout unmasks WDR33-specific contributions to complex stability and substrate recognition without genetic redundancy. This model is particularly suited to dissecting how WDR33 cooperates with CPSF1 and CPSF4 in signal recognition, and to exploring compensation mechanisms involving other RNA-binding proteins. It also offers a platform for screening small molecules that modulate 3′ end processing.

Researchers can employ the WDR33 Knockout HEK293T Cell Line in diverse experiments including RNA sequencing to map global shifts in polyadenylation site usage, co-immunoprecipitation to assess CPSF complex assembly in the absence of WDR33, and RT-qPCR or western blotting to validate downstream gene expression changes. The line is valuable for mechanistic studies of alternative polyadenylation in cancer biology and for drug screens targeting the polyadenylation machinery. It can also serve as a negative control in rescue experiments where wild-type or mutant WDR33 is re-introduced. For more information about this cell line or to request a quote, please contact Ascent Research.