The CCDC136 Knockout HEK293T Polyclonal Cells are a CRISPR/Cas9-mediated gene-disrupted polyclonal cell pool engineered to ablate expression of the coiled-coil domain-containing protein 136 (CCDC136). This polyclonal knockout population provides a heterogeneous loss-of-function model that avoids the clonal selection biases inherent to monoclonal lines, enabling robust assessment of CCDC136-dependent processes. The use of a polyclonal format ensures representation of diverse genetic alterations at the target locus, supporting broad functional studies.
HEK293T cells are a female human embryonic kidney epithelial line immortalized with sheared adenovirus 5 DNA, constitutively expressing the SV40 large T antigen. This genetic background confers exceptionally high transfection efficiency and robust recombinant protein expression, making HEK293T a preferred host for viral packaging, protein production, and functional genomics. The neutral hormonal environment of the female origin is advantageous for studies that require independence from sex-specific factors, and the cell line??s rapid growth facilitates large-scale experiments.
CCDC136 encodes a coiled-coil domain protein essential for acrosome biogenesis during spermatogenesis. The protein localizes to the acrosome, a Golgi-derived vesicular organelle, where it facilitates vesicle docking and fusion through direct interactions with GOPC. CCDC136 operates within a molecular network that includes HRB, SPATA16, ZPBP1, and DPY19L2, all critical for acrosome integrity and sperm head shaping. Disruption of this network leads to globozoospermia and male infertility. While upstream regulators of CCDC136 remain elusive, it is downstream of undefined signals and upstream of acrosome formation and sperm head morphogenesis.
Although HEK293T cells are non-spermatogenic, they provide a tractable heterologous system for studying the biochemical properties and interaction partners of CCDC136. The high transfectability enables reconstitution experiments where wild-type or mutant CCDC136 can be expressed alongside GOPC and other acrosomal proteins, allowing for co-immunoprecipitation and immunofluorescence analyses of coiled-coil domain function. This knockout background eliminates endogenous CCDC136, facilitating clean structure-function investigations and the characterization of pathogenic variants linked to male infertility.
Researchers can utilize the CCDC136 Knockout HEK293T Polyclonal Cells for validation of gene disruption via Sanger sequencing, and for quantifying CCDC136 mRNA and protein levels by RT-qPCR and Western blotting. The cells are suitable for protein?Cprotein interaction studies using co-immunoprecipitation with known partners such as GOPC, and for subcellular localization by immunofluorescence microscopy. This knockout pool also supports genome-wide CRISPR screens and transcriptomic analyses (RNA-seq) to explore compensatory pathways or off-target effects. For further technical information or custom applications, please contact Ascent Research.
