The CD47 Knockout A-549 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population derived from the human lung adenocarcinoma A-549 cell line. In this product, the CD47 gene has been disrupted using CRISPR/Cas9 technology to create a loss-of-function model. The resulting polyclonal pool comprises a heterogeneous mixture of edited cells, providing a versatile tool for studying CD47-dependent processes without requiring clonal isolation. This knockout population enables researchers to investigate the functional consequences of CD47 ablation in a well-characterized lung cancer epithelial background.
The A-549 cell line, derived from a 58-year-old male with lung adenocarcinoma, is an adherent epithelial model of alveolar Type II pulmonary epithelium. Widely used in cancer biology and drug metabolism research, A-549 cells retain features of lung adenocarcinoma, including EGFR expression and TP53 mutations. Their robust growth and genetic tractability make them suitable for CRISPR-mediated genome editing, enabling generation of knockout derivatives for mechanistic studies.
CD47 is a transmembrane immunoglobulin superfamily protein that serves as a ligand for signal regulatory protein alpha (SIRP??). CD47?CSIRP?? interaction on macrophages induces ITIM phosphorylation, recruiting SHP-1 and SHP-2 phosphatases to inhibit myosin IIA accumulation and phagocytosis. CD47 also modulates integrin signaling by associating with ??v??3 and ??IIb??3 integrins and binding thrombospondin-1, thereby regulating cell adhesion, migration, and angiogenic pathways. Downstream of CD47, PI3K/AKT/mTOR signaling promotes cell survival, while Rho GTPases Rac1 and Cdc42 mediate cytoskeletal reorganization. CD47 expression is driven by transcription factors NF-??B, STAT3, HIF1??, and MYC, and is post-transcriptionally regulated by miR-133a. Thus, CD47 integrates immune evasion with cell-intrinsic proliferative and migratory signals.
In lung adenocarcinoma, CD47 overexpression contributes to immune escape and tumor progression. A-549 cells harbor a KRAS mutation, representing a common oncogenic driver in non-small cell lung cancer. Knocking out CD47 in this background disrupts the CD47-SIRP?? immune checkpoint, enabling studies of macrophage-mediated clearance and the crosstalk between CD47 and KRAS-driven signaling. This model is thus particularly relevant for investigating how loss of CD47 affects PI3K/AKT pathway activity, integrin-mediated adhesion, and tumor cell susceptibility to innate immune attack.
This polyclonal knockout population is suited for a breadth of functional assays. It provides a negative control for CD47 detection by flow cytometry, RT-qPCR, western blotting, and immunofluorescence. The cells facilitate quantitative macrophage phagocytosis assays to evaluate CD47-dependent immune evasion and anti-CD47 therapeutic antibodies. Additionally, they support cell proliferation, migration, and invasion studies to define CD47’s role in tumor cell biology. Co-immunoprecipitation can be used to map CD47-interacting proteins. Researchers may also employ the knockout line to validate antibody specificity and explore combination therapies. For technical inquiries, please contact Ascent Research.
