The ACER1 Knockout HAP1 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal cell population derived from the HAP1 cell line, featuring targeted disruption of the ACER1 gene that encodes alkaline ceramidase 1. This polyclonal knockout product provides a heterogeneous pool of cells with loss-of-function mutations at the ACER1 locus, avoiding the need for single-cell clone isolation while retaining a high frequency of knockout alleles. The population format is ideally suited for bulk functional genomics screens, pathway interrogation, and drug sensitivity profiling, enabling robust and reproducible loss-of-function analyses in a near-haploid background.
HAP1 is a human near-haploid chronic myeloid leukemia cell line with a disomic karyotype for chromosome 8 and a segment of chromosome 15. This line retains wild-type TP53 and carries the BCR-ABL fusion kinase, the hallmark driver of chronic myeloid leukemia. The cells exhibit a fibroblast-like morphology and grow in suspension. The near-haploid genome simplifies genetic editing and reduces functional redundancy, establishing HAP1 as a prominent model for hematopoietic malignancy research, high-throughput chemical screening, and systematic functional annotation of cancer-related genes.
ACER1 encodes an alkaline ceramidase localized at the endoplasmic reticulum that hydrolyzes ceramides into sphingosine and free fatty acids, a key step in sphingolipid catabolism. By lowering ceramide levels, ACER1 indirectly promotes formation of sphingosine-1-phosphate (S1P), a bioactive lipid that signals through S1P receptors (S1PR1-5) to regulate cell survival, migration, and proliferation. ACER1 activity is counterbalanced by ceramide synthases (CERS1-6) and sphingosine kinases (SPHK1, SPHK2); the product sphingosine-1-phosphate can be further metabolized by sphingosine-1-phosphate phosphatase. Downstream, ACER1 modulates the ceramide/S1P rheostat, influencing Bcl-2 family protein-mediated mitochondrial apoptosis and autophagy pathways.
Disruption of ACER1 in the BCR-ABL-positive HAP1 background creates a powerful system to dissect the role of ceramide signaling in leukemia. Ceramide metabolism is frequently dysregulated in hematologic malignancies, where reduced ceramide and elevated S1P confer survival advantages and drug resistance. ACER1 knockout cells enable researchers to investigate how loss of ceramidase activity affects BCR-ABL downstream signaling, alters apoptotic sensitivity, and modifies responses to tyrosine kinase inhibitors such as imatinib, thereby revealing potential therapeutic vulnerabilities linked to sphingolipid homeostasis.
This polyclonal knockout model supports a broad spectrum of experimental applications, including functional genomics screens, detailed apoptosis assays using annexin V staining, cell cycle analysis by flow cytometry, and sphingolipid profiling via mass spectrometry. Researchers can quantify ceramide levels with ELISA, measure sphingosine kinase activity, monitor ACER1 protein expression by Western blotting, and assess drug sensitivity to imatinib. RNA-sequencing enables transcriptome-wide analysis of ACER1-dependent gene regulation. For further information, please contact Ascent Research.
