Cas9 Stable Cell Line - 3D4/21

The Cas9-stable 3D4/21 cell line is a genetically engineered derivative of the porcine alveolar macrophage cell line 3D4/21; the parental cell line was originally isolated from the lung tissue of domestic pigs (Sus scrofa). This modified cell line stably expresses the CRISPR-associated protein 9 (Cas9) endonuclease, thereby enabling targeted genomic editing capabilities without the need for repeated transfections. As a cell line derived from immortalized macrophages, the parental 3D4/21 cells exhibit typical phagocytic activity and express key immune markers—such as CD14 and MHC-II—making them an ideal model for porcine immunology research. This cell line demonstrates exceptionally high transfection efficiency, maintains stable Cas9 expression levels even after more than 20 passages, and retains the intrinsic biological functions of macrophages, including cytokine production and the ability to respond to pathogens.

The Cas9-stable 3D4/21 cell line offers groundbreaking application prospects in the fields of veterinary immunology and infectious disease research. With regard to porcine viral pathogens, this cell line facilitates functional genomics screening; by systematically knocking out genes involved in viral entry (e.g., CD163) or immune evasion pathways, researchers can identify host factors critical for infection by viruses such as African Swine Fever Virus (ASFV) or Porcine Reproductive and Respiratory Syndrome Virus (PRRSV). Researchers can utilize this cell line to construct macrophage-specific immune response models—by editing genes involved in inflammasome activation (e.g., NLRP3), cytokine signaling (e.g., IL-1β), or pattern recognition receptors (e.g., TLR4)—thereby enabling in-depth investigations into immunopathological mechanisms. In the realm of vaccine development, this cell line can be used to enhance vaccine immunogenicity by generating knockout strains targeting interferon-suppressing genes, thereby accelerating the development of vaccines based on reverse genetics strategies.