This product comprises a polyclonal population of HAP1 cells in which the ANGEL2 gene has been disrupted using CRISPR/Cas9-mediated gene editing. ANGEL2 encoding a 2′,3′-cyclic nucleotide 3′-phosphodiesterase plays an essential role in tRNA splicing by removing the 2′,3′-cyclic phosphate at splice junctions, a critical step for subsequent ligation and maturation of intron-containing tRNAs. The resulting polyclonal knockout cells provide a heterogeneous loss-of-function model suitable for studying ANGEL2-dependent processes without clonal artifacts.
The HAP1 cell line is a near-haploid, fibroblast-like cell line originally derived from the KBM-7 chronic myeloid leukemia line from a male patient. Its haploid karyotype makes it exceptionally valuable for genetic screens, gene-trap mutagenesis, and functional genomics studies. The cells grow adherently and maintain a stable near-haploid state, facilitating the isolation and characterization of gene disruptions. HAP1 cells retain key signaling pathways relevant to cancer biology and stress responses, offering a simplified genomic context for dissecting gene function.
ANGEL2 functions as a 2′,3′-cyclic nucleotide 3′-phosphodiesterase that specifically hydrolyzes the 2′,3′-cyclic phosphate group generated at tRNA intron splice junctions. This activity directly produces 3′-phosphate and 2′-hydroxyl ends on exon halves, permitting proper ligation by the RTCB-containing tRNA ligase complex, which also includes cofactors CGI-99 and FAM98B. ANGEL2 expression is regulated by the unfolded protein response (UPR) transcription factors ATF4 and XBP1, linking its activity to cellular stress signaling. Downstream, ANGEL2-mediated tRNA maturation is essential for efficient translation elongation and global protein synthesis, particularly under conditions of ER stress. Disruption of ANGEL2 impairs the production of mature functional tRNAs, leading to translational defects and potentially activating the UPR through feedback mechanisms.
In the HAP1 background, ANGEL2 knockout cells provide a powerful tool to investigate tRNA processing within a haploid genetic system. The near-haploid state facilitates unambiguous assignment of phenotypes to ANGEL2 loss, as there is no diploid compensation. This model is particularly suited for integrated studies combining genetic screens with biochemical assays, allowing researchers to dissect ANGEL2-dependent steps in tRNA splicing and translation regulation. Moreover, the CML origin of HAP1 cells offers a relevant setting for exploring ANGEL2’s potential roles in leukemia pathogenesis, where altered tRNA metabolism and translational control may contribute to disease progression and drug resistance.
The ANGEL2 Knockout HAP1 Polyclonal Cells support haploid genetic screens, tRNA processing studies, and disease modeling. Specific assays include western blotting for UPR markers, RT-qPCR for tRNA splicing intermediates, and polysome profiling for translation efficiency. Researchers can investigate neurodevelopmental disorders such as microcephaly with seizures and spastic paraplegia, as well as leukemia, where ANGEL2 mutations or dysregulation are implicated. Drug target validation under ER stress conditions and RNA-seq-based transcriptomics are additional key applications. For detailed technical inquiries or to discuss custom experimental applications, please contact Ascent Research.
