KHDRBS1 Knockout HEK293T Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population derived from HEK293T cells, designed for loss-of-function studies of the KHDRBS1 gene. The polyclonal format provides a diverse mixture of edited alleles, enabling robust analysis of gene disruption effects without clonal selection bias. This target-gene disruption model is produced using CRISPR/Cas9-mediated genome editing to introduce targeted mutations in KHDRBS1, resulting in a heterogeneous population suitable for pooled functional genomics and biochemical assays.
The host cell line, HEK293T, is an adherent human embryonic kidney epithelial line expressing the SV40 large T antigen, which supports episomal replication of plasmids containing the SV40 origin. This feature makes HEK293T cells a workhorse for protein expression, lentivirus production, and gene editing experiments. Their high transfection efficiency and robust growth enable straightforward delivery of CRISPR components and subsequent expansion of polyclonal knockout populations for downstream applications.
KHDRBS1 (Sam68) is an RNA-binding protein that integrates signals from tyrosine kinases and the mTOR pathway to coordinate alternative splicing, mRNA transport, and translation. It is activated by SRC family kinases such as FYN, and functions downstream of growth factor receptors including EGFR and the insulin receptor. Upon phosphorylation, KHDRBS1 modulates splicing of key targets like CD44, BCL2L1 (Bcl-xL), and CCND1 (cyclin D1), thereby influencing cell cycle progression and apoptosis. It interacts with signaling proteins PLCG1, GRB2, PIK3R1, and splicing regulators like HNRNPA1, linking signal transduction to post-transcriptional gene regulation. Through these interactions, KHDRBS1 serves as a node connecting mTOR, MAPK/ERK, and SRC pathways to alternative splicing decisions that control cell fate.
In HEK293T cells, which possess an active signaling milieu and are frequently used to study cancer-related pathways, disruption of KHDRBS1 removes a critical mediator of splicing-dependent survival and proliferation cues. The knockout model allows researchers to dissect how loss of this adaptor protein alters CD44 isoform switching, BCL2L1 splicing (producing pro-apoptotic Bcl-xS versus anti-apoptotic Bcl-xL), and expression of downstream effectors like IL6. This system is particularly valuable for elucidating how oncogenic kinases (e.g., SRC, FYN) couple to RNA processing events, and for probing mTOR-dependent regulation of splicing independent of translational control.
Typical uses include genome-wide splicing analysis via RNA-seq to identify KHDRBS1-dependent exon usage, paired with co-immunoprecipitation of SRC-FYN complexes to study kinase?Cadaptor interactions. The polyclonal population supports cell proliferation and apoptosis assays (e.g., flow cytometry for annexin V) to quantify functional outcomes of splicing dysregulation. Researchers can perform RT-qPCR to validate isoform shifts in CD44v and Bcl-xL, and Western blotting to confirm loss of KHDRBS1 and altered phospho-tyrosine signaling. These applications make the product suitable for studies in cancer biology, signal transduction, and drug resistance mechanisms, particularly in the context of mTOR or MAPK pathway-driven malignancies. For additional details or custom requirements, please contact Ascent Research.
