The HNRNPLL Knockout HEK293T Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population generated from the HEK293T human embryonic kidney line. This product carries a targeted disruption of HNRNPLL, creating a loss-of-function model for studying the RNA-binding protein??s role in alternative splicing. HNRNPLL controls exon inclusion by binding to exonic splicing silencers; its knockout here facilitates dissection of splicing network changes without clonal selection. The polyclonal population encompasses a variety of edited alleles, providing a heterogeneous pool that can be directly used for population-based functional assays such as RNA-seq or pooled screens, without single-cell cloning required.
HEK293T is a human embryonic kidney epithelial cell line transformed with adenovirus 5 DNA and stably expressing SV40 large T antigen, which drives high-copy plasmid replication and boosts transfection efficiency. Originally established by sheared adenovirus 5 DNA transformation, HEK293T cells are a subclone of HEK293 that stably express the SV40 large T antigen, which enhances episomal replication from SV40 ori-bearing plasmids. This leads to high transgene expression levels and efficient transient transfection, making them a workhorse for mammalian cell biology and CRISPR-based gene editing. Widely used for recombinant protein expression, viral vector production, and CRISPR screening, HEK293T offers rapid growth and a well-characterized genomic background, providing a robust platform for gene regulation studies.
HNRNPLL is an hnRNP-family splicing factor that interacts with HNRNPL, SRSF proteins, and the spliceosome. In T cells, TCR stimulation upregulates HNRNPLL via NFATc2 and NF-??B. It then targets CD45 (PTPRC) pre-mRNA, promoting exon 4?C6 skipping to switch isoform expression from CD45RO to CD45RA. This alters TCR signaling strength and tunes immune responses. HNRNPLL belongs to the heterogeneous nuclear ribonucleoprotein (hnRNP) family and acts as a splicing regulator by binding to exonic splicing silencers and forming complexes with HNRNPL, SRSF proteins, and core spliceosomal components, thereby modulating the threshold for T cell activation.
Although HEK293T cells lack the full T cell signaling apparatus, they express core splicing components, offering a simplified context to probe HNRNPLL??s intrinsic splicing function. This model is ideal for dissecting HNRNPLL??s RNA-binding specificity, its interplay with partner proteins, and its impact on constitutive or alternative splicing of both endogenous genes and introduced minigene substrates. The polyclonal knockout pool can be paired with minigene reporters or used to examine endogenous splicing events, enabling biochemical analysis of RNA-binding and protein interactions independent of immune-cell signaling. The absence of immune-specific signals allows focused study of splicing mechanisms.
Key applications include RNA-seq to globally profile alternative splicing changes, targeted RT-PCR and flow cytometry for CD45RA/RO isoform analysis (when relevant reporters are introduced), minigene splicing reporter assays to define regulatory elements, RNA immunoprecipitation (RIP) to identify direct RNA targets, and co-immunoprecipitation to characterize protein?Cprotein interactions. The cells can also be integrated into pooled or arrayed CRISPR screens aimed at identifying novel splicing regulators in autoimmune disease pathways. For further details or technical support, please contact Ascent Research.
