The CD27 Knockout HAP1 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population derived from the haploid human HAP1 cell line, targeting CD27. This loss-of-function model is generated via CRISPR/Cas9-mediated gene disruption, yielding a heterogeneous pool of edited cells that collectively eliminate CD27 expression. As a polyclonal reagent, it enables studies of CD27-dependent processes without clonal isolation, while maintaining the parental line??s genetic integrity at non-targeted sites.
HAP1 is a haploid human cell line originally derived from KBM-7 chronic myeloid leukemia cells, prized for genetic research due to its single-copy genome that simplifies editing outcomes. The haploid state ensures that disruption of the one CD27 allele directly results in complete loss of function, avoiding heterozygous effects. Its robust growth and standard culture compatibility make HAP1 a suitable host for CRISPR knockout investigations in signaling and immune research.
CD27 encodes a costimulatory immune receptor of the TNF receptor superfamily, essential for T-cell activation, proliferation, survival, and B-cell differentiation. Ligand CD70 binding triggers recruitment of TRAF2 and TRAF5, activating the IKK complex and downstream NF-??B, while also stimulating the JNK cascade and PI3K-Akt pathway. These signals upregulate Bcl-xL, Cyclin D1, and IL-2, promoting lymphocyte expansion. CD27 also interacts with SIVA1, which may modulate apoptotic responses.
In HAP1 cells, CD27 knockout eliminates CD70-induced NF-??B and JNK activation, abrogating pro-survival transcriptional outputs. Despite not being a lymphocyte, HAP1??s hematopoietic origin retains signaling components, enabling reconstitution with relevant ligands or effectors to dissect CD27 pathways. The haploid background ensures unambiguous phenotype interpretation by removing wild-type allele interference.
Applications include immune cell signaling studies, costimulatory receptor function analysis, cancer immunotherapy target validation, and autoimmune disease research. Typical assays involve western blotting, flow cytometry, NF-??B reporter assays, phospho-JNK ELISA, apoptosis tests upon CD70 stimulation, TRAF2 co-immunoprecipitation, and cytokine secretion profiling. For further information or custom application inquiries, contact Ascent Research.
