The CD177 Knockout HAP1 Polyclonal Cells constitute a CRISPR/Cas9-edited polyclonal cell population carrying targeted disruption of the CD177 gene in the HAP1 human haploid cell line. This polyclonal knockout pool offers a heterogeneous loss-of-function model, allowing robust functional interrogation of CD177 biology without clonal artefacts. The CRISPR-mediated gene disruption delivers a stable, heritable knockout background suitable for diverse biochemical and cell-based assays.
The HAP1 host line is derived from the KBM-7 chronic myeloid leukemia cell line and maintains a near-haploid karyotype, making it a versatile platform for functional genomics and haploid genetic screens. Its fibroblast-like adherent growth and genetic tractability facilitate high-throughput gene editing and phenotypic analysis. Haploidy enables complete gene inactivation via disruption of a single allele, minimizing compensatory effects and simplifying genotype?Cphenotype correlations.
CD177 is a glycosylphosphatidylinositol-anchored glycoprotein selectively expressed on neutrophils, where it critically regulates transendothelial migration. Mechanistically, CD177 engages endothelial PECAM-1 (CD31), triggering inside-out activation of ??2 integrins (LFA-1 and Mac-1), which promotes firm adhesion to ICAM-1 and subsequent extravasation. CD177 expression is induced by G-CSF receptor signaling and the myeloid transcription factors C/EBP?? and PU.1, and is further modulated by inflammatory cytokines such as TNF-?? and IL-1??. Downstream, CD177-mediated signals activate Rho GTPases, leading to cytoskeletal rearrangement and enhanced neutrophil adhesion and motility. The protein also interacts with Proteinase 3 (PRTN3) and indirectly associates with ??2 integrins, placing it at a central node in leukocyte adhesion and inflammatory cascades.
Integrating the CD177 knockout into the HAP1 background creates a genetically simplified system to dissect the receptor’s molecular functions, even though HAP1 cells are not of hematopoietic origin. Ectopic expression of pathway components or cell-free interaction setups allows detailed study of CD177-dependent adhesion and signaling. The polyclonal format preserves natural mutation diversity, reducing bias from individual clones and enhancing the detection of robust biological effects. Combined with the haploid state, this model provides unambiguous links between gene disruption and observed phenotypes, supporting high-content screening and mapping of genetic interaction networks.
This product is suitable for investigations into CD177-PECAM-1 binding dynamics, regulation of ??2 integrin activation, and the mechanics of neutrophil migration. Typical assays include Sanger sequencing for knockout confirmation, Western blotting, flow cytometric analysis of surface markers, and solid-phase adhesion assays using recombinant PECAM-1. Transendothelial migration models and co-immunoprecipitation further expand the utility to probe protein?Cprotein interactions and signal transduction. For further information on product usage and customization, please contact Ascent Research.
