The CCDC137 Knockout HEK293T Polyclonal Cells product comprises a heterogeneous population of HEK293T cells subjected to CRISPR/Cas9-mediated gene disruption targeting the CCDC137 locus. This polyclonal knockout pool provides a robust loss-of-function model for investigating nucleolar biology without the confounding effects of clonal selection. The use of polyclonal cells preserves genetic diversity, mitigating clonal artifacts and enabling more representative studies of CCDC137 function.
HEK293T cells are immortalized human embryonic kidney derivatives stably expressing SV40 large T-antigen, conferring high transfection efficiency and episomal replication of plasmids with SV40 origin. These cells are widely employed in protein expression, viral production, and genomic perturbation studies due to their robust growth and ease of manipulation. In the context of CCDC137 knockout, HEK293T provides a permissive background for dissecting nucleolar functions and ribosome biogenesis pathways.
CCDC137 encodes a nucleolar protein implicated in ribosome biogenesis, specifically in pre-rRNA processing and 18S rRNA maturation. The protein interacts with key nucleolar factors including nucleophosmin, fibrillarin, and nucleolin, forming part of the machinery that processes ribosomal RNA precursors. CCDC137 expression is regulated by upstream oncogenic signals, notably MYC and mTOR, which coordinate ribosome production with cellular growth demands. Consequently, disruption of CCDC137 likely impairs pre-rRNA cleavage steps, leading to defective 40S ribosomal subunit assembly, diminished protein synthetic capacity, and attenuated cell proliferation. This positions CCDC137 at a critical node linking nutrient and growth factor signaling to ribosome biogenesis.
In HEK293T cells, a rapid proliferative background with active ribosome biogenesis, loss of CCDC137 is expected to reveal specific vulnerabilities in nucleolar stress responses and translational control. The polyclonal knockout pool enables analysis of heterogeneous cellular outcomes, recapitulating the stochastic nature of gene disruption akin to primary cell studies. This model is particularly relevant for cancer biology, as CCDC137 has been implicated in hepatocellular carcinoma and other malignancies where ribosome biogenesis is dysregulated. Investigations using these cells can elucidate how MYC/mTOR-driven ribosomal overproduction depends on individual nucleolar cofactors.
Typical applications include kinetic monitoring of pre-rRNA processing intermediates by RT-qPCR, assessment of nucleolar morphology via immunofluorescence for fibrillarin or nucleophosmin, polysome profiling to evaluate translation efficiency, and cell proliferation assays to quantify growth defects. Western blotting can confirm CCDC137 downregulation and assess levels of interacting partners or markers of ribosome assembly stress. These cells also serve as a tool for drug discovery screens targeting the ribosome biogenesis pathway. For further technical assistance, please contact Ascent Research.
