The CD300A Knockout HAP1 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population designed to disrupt the CD300A gene in the HAP1 cell line. This product provides a heterogeneous pool of edited cells, each carrying distinct disruption events at the target locus, enabling loss-of-function studies without the need for single-cell cloning. The CRISPR/Cas9-mediated gene disruption generates a knockout model suitable for investigating the functional roles of CD300A in immune regulation, signal transduction, and disease contexts.
The HAP1 cell line is a near-haploid human chronic myeloid leukemia (CML) cell line derived from the KBM-7 line, characterized by its haploid karyotype for most chromosomes except a disomic region on chromosome 15. This genetic minimalism minimizes functional redundancy and facilitates unambiguous genotype-phenotype correlations, making HAP1 an ideal host for CRISPR-based knockouts. As a CML model, HAP1 retains key signaling pathways relevant to myeloid biology and immune interactions, providing a physiologically relevant platform for studying immune receptor function.
CD300A is an ITIM-bearing inhibitory immune receptor that recognizes phosphatidylserine exposed on apoptotic cells, a process triggered by upstream cellular stress signals. Upon ligand engagement, CD300A is phosphorylated by Src family kinases such as Lyn, leading to the recruitment and activation of the tyrosine phosphatases SHP-1 and SHP-2 through its ITIM motifs. These phosphatases dephosphorylate downstream targets, inhibiting NF-??B and MAPK signaling pathways. CD300A interacts with the Fc??RI?? chain, modulating immunoreceptor signaling as a negative regulator of immune activation.
In CML, CD300A-mediated inhibitory signaling is relevant for understanding immune evasion and inflammatory regulation. The HAP1 knockout model enables precise dissection of CD300A’s role in modulating myeloid cell function, cytokine secretion, and interactions with apoptotic cells. The near-haploid genome ensures that phenotypes are clearly attributable to CD300A disruption, minimizing compensatory effects. This model is valuable for exploring CD300A in autoimmune diseases, inflammatory disorders, and cancer immune evasion where inhibitory receptor signaling is deregulated.
Researchers can employ these polyclonal knockout cells in a variety of advanced applications, including immune checkpoint research, inhibitory receptor signaling studies, functional genomics screens, and drug target validation. Representative assays include Western blotting to assess CD300A protein levels, flow cytometry to monitor cell-surface receptor expression, cytokine secretion assays to measure inflammatory outputs, phospho-signaling analysis to evaluate NF-??B and MAPK pathway activity, and co-culture experiments with apoptotic cells to examine ligand-induced signaling. For further details or technical support, please contact Ascent Research.
