The CCNL2 Knockout HAP1 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population designed for loss-of-function studies of the CCNL2 gene in a haploid human cell background. This polyclonal knockout model comprises a heterogeneous pool of HAP1 cells with disruption of the CCNL2 locus, enabling functional genomics research without clonal expansion. It offers a versatile tool for investigating CCNL2-mediated processes in transcription and splicing regulation.
HAP1 is a haploid human cell line derived from the KBM-7 chronic myeloid leukemia (CML) line. Its near-haploid karyotype greatly facilitates gene editing, as disruption of a single allele is sufficient to generate complete knockout phenotypes, making it a workhorse for CRISPR-based functional genomics screens and arrayed genetic perturbations. The hematopoietic origin of HAP1 cells provides a contextually relevant model for studying genes involved in blood cancers and for dissecting cell cycle regulatory networks that are often subverted in leukemia.
CCNL2 encodes cyclin L2, a regulatory cyclin that forms an active kinase complex with CDK11. This complex directly phosphorylates the C-terminal domain of RNA polymerase II (POLR2A) and serine/arginine-rich splicing factors such as SRSF1, thereby coordinating transcription elongation with pre-mRNA splicing. Through this phosphorylation, CCNL2 integrates transcriptional activity with post-transcriptional processing, a control mechanism that is essential for proper gene expression and is frequently deregulated in cancer. CCNL2 also intersects with cell cycle pathways, further underscoring its significance in oncogenic processes.
In the haploid HAP1 knockout setting, disruption of CCNL2 permits precise functional analysis of its role in transcription-coupled splicing, unambiguously linking phenotypic outcomes to gene loss. This model enables the dissection of how CDK11-dependent phosphorylation by CCNL2 influences RNA polymerase II processivity and splicing factor activation. Given the CML origin of HAP1 cells, the knockout system is particularly relevant for investigating splicing abnormalities that may promote leukemic transformation and for evaluating CCNL2 as a vulnerability in cancer cells.
Applications include transcriptome-wide profiling by RNA-seq to identify splicing signatures dependent on CCNL2, co-immunoprecipitation to probe interactions with CDK11 and splicing regulators, and Western blotting or RT-qPCR to quantify expression changes in downstream effectors like POLR2A and SRSF1. Immunofluorescence can be used to examine subcellular localization of splicing factors. This product is also amenable to high-content screening for compounds that modulate splicing. For additional assistance, please contact Ascent Research.
