The HNRNPF Knockout HEK293T Polyclonal Cells represent a CRISPR/Cas9-edited polyclonal knockout population engineered to disrupt the HNRNPF gene in the human HEK293T cell line. This polyclonal cell product provides a loss-of-function model for studying the regulatory roles of the HNRNPF RNA-binding protein in alternative splicing, mRNA stability, and translation. By eliminating functional HNRNPF expression across a heterogeneous cell population, researchers can investigate global splicing alterations and downstream cellular effects without the biases inherent in single-clone isolation.
The host cell line, HEK293T, is a widely utilized human embryonic kidney epithelial line derived from HEK293 cells that stably express the SV40 large T antigen. This modification enables robust transient expression and efficient lentiviral and retroviral vector production, making HEK293T optimal for functional genomics. Originally transformed with sheared adenovirus type 5 DNA, these cells provide a transfectable platform for knockout studies.
HNRNPF functions as a key splicing regulator by binding guanosine-rich motifs within pre-mRNAs, influencing spliceosome assembly and exon selection. Its activity is modulated by SR protein kinases SRPK1 and SRPK2, which phosphorylate serine/arginine-rich factors, and by cellular stress signals. HNRNPF interacts with core spliceosome components like SF3B, U2AF2, and RNA polymerase II CTD, and cooperates with hnRNP H and hnRNP A1. It regulates alternative splicing of targets such as Bcl-x, Grin1, CD44, and E-cadherin, affecting apoptosis, neuronal receptor function, and cell adhesion.
In the HEK293T background, HNRNPF knockout enables dissection of its splicing functions independent of tissue-specific factors. The epithelial origin makes it relevant for studying epithelial-mesenchymal transition via CD44 and E-cadherin isoforms and for investigating apoptotic gene regulation in cancer. The polyclonal nature reduces clonal artifacts, offering a heterogeneous model aligned with tumor biology and neuropathological conditions like glioblastoma and ALS.
This knockout pool supports RNA-seq and CLIP-seq to map HNRNPF-dependent splicing events, quantitative RT-PCR and minigene assays for isoform validation, and apoptosis assays linking splicing to cell death. It facilitates high-throughput screening of splicing modulators for drug discovery and advances research into lung cancer, breast cancer, and splicing-related neuropathologies such as amyotrophic lateral sclerosis. For ordering information, technical support, or custom projects, please contact Ascent Research.
