The C3orf49 Knockout HEK293T Polyclonal Cells are a CRISPR/Cas9-edited polyclonal population derived from HEK293T human embryonic kidney cells, engineered for targeted disruption of the C3orf49 gene. This heterogeneous knockout model is produced by introducing Cas9 and guide RNAs against C3orf49, yielding a mixture of cells with diverse edits. The polyclonal format maintains genetic diversity and provides a loss-of-function tool for studying C3orf49. It is optimally suited for high-throughput screening and functional genomics without requiring clonal isolation, facilitating examination of its role in cell cycle and apoptosis.
The HEK293T parental line is a widely used derivative of HEK293 cells that stably expresses SV40 large T antigen, enhancing episomal plasmid replication and recombinant protein yield. These cells are central to biomedical research for transient expression, lentiviral packaging, and as a tractable cancer model. Their rapid proliferation, high transfection efficiency, and compatibility with diverse assays render them an excellent system for gene knockout. The C3orf49 knockout in this background permits direct functional interrogation within a context that mimics aspects of deregulated growth.
C3orf49 is a putative modulator of cell proliferation and apoptosis, linked to the p53 pathway. It is transcriptionally controlled by E2F factors and p53, ensuring cell cycle?Cdependent expression. Disruption of C3orf49 alters downstream effectors cyclin D1 and CDK4, which govern G1/S progression, and shifts the BAX/BCL2 apoptotic balance. Although direct interactors remain elusive, C3orf49 likely integrates signals within the p53?CBAX?CBCL2?Ccyclin D1?CCDK4 network. Its loss is predicted to dysregulate proliferation and apoptotic thresholds, potentially fueling oncogenic processes.
In HEK293T cells, where SV40 large T antigen inactivates p53 and Rb, the C3orf49 knockout offers a setting to probe how this gene interfaces with residual proliferation and survival pathways. This model facilitates dissection of C3orf49??s role in modulating apoptosis sensitivity and G1/S transition under compromised tumor suppression. Comparative analyses with parental cells allow identification of C3orf49-dependent changes in cell cycle kinetics and apoptotic responses, with implications for glioblastoma and hepatocellular carcinoma, where C3orf49 has been associated.
Researchers employ these cells in quantitative proliferation assays (MTT, BrdU), Annexin V flow cytometry for apoptosis, and immunoblotting for cyclin D1, CDK4, BAX, and BCL2. The polyclonal population supports drug screening, RNA-seq, and proteomic studies to elucidate C3orf49-linked networks. This model provides statistical power for phenotypic variability analysis and is a valuable tool for cancer biology and preclinical testing of p53 pathway modulators. For further information, please contact Ascent Research.
