The ELAVL2 Knockout HAP1 Polyclonal Cells consist of a CRISPR/Cas9-edited polyclonal population of HAP1 cells with disrupted ELAVL2. This pooled knockout model avoids clonal artifacts, providing a robust system for functional studies. The polyclonal format ensures genetic diversity and consistent target disruption, making it suitable for population-based assays and high-throughput screening applications.
HAP1 is a near-haploid human fibroblast-like cell line derived from the KBM-7 chronic myelogenous leukemia line. Its single-copy genome allows complete gene knockout via targeting one allele, eliminating heterozygosity. With adherent morphology and active signaling pathways, HAP1 is widely used for genome-wide loss-of-function screens and mechanistic investigations across various fields.
ELAVL2 is a neuron-specific RNA-binding protein that recognizes AU-rich elements in the 3??UTRs of target mRNAs. It stabilizes transcripts such as GAP-43, MAP2, Tau, and c-fos, and its expression is driven by transcription factors NEUROD1 and ASCL1, with modulation by BDNF signaling. ELAVL2 interacts with other ELAV family members (HuR, HuC, HuD) and poly(A)-binding protein to form ribonucleoprotein granules that are transported by kinesins. This post-transcriptional network governs the spatiotemporal expression of proteins required for neuronal differentiation, axon pathfinding, and synaptic plasticity. The knockout in HAP1 provides a clean genetic background to study these RNA regulatory mechanisms.
Although HAP1 cells are non-neuronal, the ELAVL2 knockout offers a simplified system to dissect core ARE-dependent regulatory functions. The near-haploid genome ensures unambiguous genotype-phenotype relationships, enabling precise analysis of mRNA stability and translation. The model also serves as a negative control for antibody specificity and allows structure-function rescue experiments. Interaction studies with the ubiquitous HuR protein can uncover differences in ARE-binding protein network dynamics.
Applications include actinomycin D chase and luciferase reporter assays to quantify mRNA decay and translational regulation, and RNA-seq for transcriptome-wide target identification. The cells support high-throughput screens for small-molecule modulators of ARE-dependent pathways and pooled CRISPR genetic modifier screens. Biochemical techniques such as RNA immunoprecipitation and Western blotting further enable mechanistic studies. This knockout model is a valuable tool for drug discovery in neurodegeneration and paraneoplastic disorders. For further information, contact Ascent Research.
