The BST1 Knockout HAP1 Polyclonal Cells product consists of a CRISPR/Cas9-edited polyclonal knockout cell population in the human HAP1 near-haploid cell line, engineered to disrupt the BST1 gene. This polyclonal population provides a heterogeneous pool of loss-of-function alleles suitable for pooled screening and functional studies without single-cell cloning artifacts. The knockout model enables robust investigation of BST1-dependent phenotypes in a well-characterized genetic background, supporting applications in cancer biology, immunology, and signal transduction research.
HAP1 cells are a near-haploid human line derived from KBM-7 chronic myeloid leukemia (CML) cells, retaining a single copy of most chromosomes. This ploidy facilitates efficient gene disruption, making HAP1 a popular model for CRISPR screens. They express myeloid lineage markers and maintain BCR-ABL1-driven signaling, while their adherent growth supports high-throughput and imaging assays.
BST1 encodes CD157, a GPI-anchored ectoenzyme catalyzing the conversion of NAD+ to cyclic ADP-ribose (cADPR), which mobilizes intracellular calcium through ryanodine and IP3 receptors. It also acts as an adhesion molecule, binding fibronectin and integrins to facilitate immune cell migration. Its expression is regulated by fibronectin, TNF-??, and IL-6, and its downstream effectors include NF-??B, FAK, and calcium-sensitive transcription factors. CD157 cooperates with CD38 in NAD+ metabolism, linking metabolic sensing to integrin-mediated adhesion and transcriptional control.
In HAP1 CML-derived cells, BST1 knockout dissects the interplay between NAD+ metabolism, calcium signaling, and adhesion in leukemic contexts. CML cells rely on altered adhesion and migration for disease progression; CD157 contributes to hematopoietic cell trafficking. Loss of BST1 enables examination of integrin-mediated attachment, fibronectin signaling, and NF-??B activity, elucidating mechanisms of leukemic survival. The near-haploid background minimizes allele redundancy, yielding clear phenotypic readouts.
This BST1 knockout model is suited for diverse assays: Western blotting and flow cytometry to assess CD157 protein loss, NAD+-metabolism assays to measure ADP-ribosyl cyclase activity, and calcium mobilization assays using Fluo-4 or Fura-2. Cell adhesion and migration on fibronectin substrates probe integrin functions; RT-qPCR and RNA-seq enable transcriptomic profiling post-disruption. The polyclonal population excels in functional genomics screens, drug target validation for immune disorders, and Parkinson’s disease research. For further information, contact Ascent Research.
