The KLHL35 Knockout HeLa Polyclonal Cells represent a CRISPR/Cas9-edited polyclonal knockout cell population in which the human KLHL35 gene has been disrupted, providing a loss-of-function model for functional studies. This polyclonal configuration comprises a pooled population of cells carrying diverse CRISPR/Cas9-mediated gene disruptions, avoiding clonal selection biases and enabling robust population-level analyses. The heterogeneous composition ensures broad representation of knockout events, making it a versatile tool for investigating the biological roles of KLHL35 in a cancer cell background.
HeLa cells, a cervical epithelial cell line derived from an HPV18-positive adenocarcinoma, serve as the host for this knockout model. These cells are among the most widely used human cancer cell lines in biomedical research, particularly suited for studying oncogenic signaling, viral transformation mechanisms, and cellular stress responses. The HPV18-positive status of HeLa cells provides a relevant context for examining how protein degradation pathways contribute to cervical cancer biology and the maintenance of the transformed phenotype.
The KLHL35 gene encodes a substrate-specific adaptor protein for the Cullin3-RING E3 ubiquitin ligase (CRL3) complex. KLHL35 directly interacts with the scaffold protein CUL3 and the RING-domain protein RBX1, forming an active E3 ligase that recruits target substrates for polyubiquitination. These ubiquitinated proteins are then directed to the 26S proteasome for degradation. Through this mechanism, KLHL35 functions as a critical regulator of protein homeostasis, mediating the turnover of yet-unidentified substrates. While upstream regulatory signals and specific downstream targets of KLHL35 remain largely uncharacterized, its established interactions with CUL3 and RBX1 place it firmly within the ubiquitin-proteasome system, a pathway essential for controlling protein abundance, cell cycle progression, and apoptosis.
Disruption of KLHL35 in HeLa cells enables the systematic interrogation of CRL3-dependent ubiquitination in a cancer-relevant epithelial model. This knockout system can be used to identify downstream substrates whose stability is governed by KLHL35, as well as to assess the phenotypic consequences of impaired ubiquitin-mediated degradation. Given the HPV-transformed nature of HeLa cells, loss of KLHL35 may reveal specific vulnerabilities or adaptive responses tied to protein degradation defects, thereby informing potential therapeutic strategies for cervical adenocarcinoma or other protein degradation disorders.
These polyclonal knockout cells are well-suited for a wide range of experimental applications, including ubiquitination pathway analysis, protein interaction studies via co-immunoprecipitation, and functional genomics approaches such as RNA-seq. Researchers can employ western blotting, ubiquitination assays, cell proliferation assays, and apoptosis assays to characterize the functional impact of KLHL35 loss. The model supports investigations into cancer biology and diseases associated with protein degradation. For further technical details or support, please contact Ascent Research.
