The KHDRBS1 Knockout HCT 116 Polyclonal Cells represent a CRISPR/Cas9-edited polyclonal knockout cell population derived from the HCT 116 human colorectal carcinoma cell line, featuring targeted disruption of the KHDRBS1 gene (also known as Sam68). This heterogeneous pool of knockout cells is generated through CRISPR/Cas9-mediated gene editing, resulting in a population-wide loss of functional KHDRBS1 protein. The polyclonal format offers a practical and efficient approach for investigating KHDRBS1-dependent cellular processes without the clonal selection bottlenecks, making it suitable for bulk functional assays and pathway analysis.
The parental HCT 116 cell line is a widely used epithelial colorectal carcinoma model, derived from a male patient. It harbors a KRAS G13D mutation and is mismatch repair proficient, ensuring genetic stability. These cells are well-characterized for studies on proliferation, DNA repair, and drug responses, and they support xenograft tumor formation.
KHDRBS1/Sam68 is an RNA-binding protein that integrates inputs from key signaling cascades to regulate alternative splicing, mRNA processing, and translation. It is phosphorylated and activated by ERK1/2 (MAPK3/MAPK1) downstream of MAPK/ERK signaling and by Src family kinases in response to insulin/IGF-1 stimulation and genotoxic stress. Once activated, Sam68 modulates the alternative splicing of BCL2L1 (BCL-X) and CD44, generating pro-survival BCL-XL or pro-apoptotic BCL-XS isoforms and CD44 variant isoforms associated with metastasis. Sam68 interacts with RNA polymerase II, PRMT1, and hnRNP proteins, and influences the expression of CCND1 (cyclin D1) and SRSF1, thereby linking extracellular cues to cell cycle progression and apoptosis. Thus, Sam68 serves as a critical node where ERK and SRC signaling converge to control splicing-dependent cell fate decisions.
In HCT 116 colorectal carcinoma cells, KHDRBS1 knockout provides a powerful tool to dissect the role of alternative splicing in cancer biology. Given the KRAS-mutant background and active MAPK/ERK and PI3K/AKT pathways, ablation of Sam68 allows researchers to examine how splicing-dependent mechanisms contribute to tumor cell proliferation, apoptosis resistance, and drug sensitivity. This model is particularly relevant for studying the interplay between oncogenic signaling and post-transcriptional gene regulation, as demonstrated by altered BCL-X isoform ratios and CD44 splicing patterns. Furthermore, the polyclonal knockout pool enables the assessment of heterogeneous cellular responses, mimicking the diversity found in tumor populations.
This knockout product is well-suited for a broad range of experimental applications, including alternative splicing analysis via RT-qPCR or RNA-seq to quantify isoform shifts in targets such as BCL-X and CD44, functional assays measuring cell proliferation and apoptosis, and drug sensitivity screening to evaluate Sam68-dependent chemoresistance. Colony formation and xenograft tumor models can be employed to assess tumorigenic potential in vivo. Additionally, the cells can be used for mechanistic studies using Western blotting to monitor signaling pathway activity and for functional genomics screens. For more information or to discuss custom applications, please contact Ascent Research.
