The JAGN1 Knockout HEK293T Polyclonal Cells product provides a heterogenous CRISPR/Cas9-edited knockout cell pool targeting the JAGN1 gene in the HEK293T human embryonic kidney cell line. JAGN1 encodes Jagunal homolog 1, an endoplasmic reticulum membrane protein essential for neutrophil differentiation and survival. The polyclonal knockout format preserves genetic diversity within the population, making it suitable for pooled functional studies and minimizing clonal artifacts.
HEK293T cells are a versatile host background, constitutively expressing the SV40 large T antigen for episomal plasmid replication and high-titer protein production. These adherent epithelial cells are easily transfectable and widely used in gene editing, signaling, and virology studies. Their robust expression machinery and well-mapped stress pathways allow detailed investigation of JAGN1??s role in ER membrane organization and homeostasis.
At the molecular level, JAGN1 interacts with reticulon proteins and NGLY1 to shape and stabilize the ER network. Loss of JAGN1 triggers ER stress, activating the unfolded protein response through the transmembrane sensors IRE1??, PERK, and ATF6. This cascade upregulates transcription factors ATF4 and CHOP, the chaperone GRP78/BIP, and the spliced form of XBP1. In the context of granulopoiesis, JAGN1 acts downstream of G-CSF signaling via the CSF3R receptor and the JAK-STAT pathway, where STAT3 activation leads to induction of CEBPA and ELF4 transcription factors. Disruption of this pathway due to JAGN1 deficiency results in impaired neutrophil granulocyte differentiation and increased apoptosis, recapitulating features of severe congenital neutropenia.
Although HEK293T cells are non-hematopoietic, they provide a clean cellular system to dissect the ER-intrinsic functions of JAGN1 without confounding differentiation signals. The cells express key UPR mediators and can be challenged with ER stressors to examine signaling kinetics and downstream apoptosis. This model enables focused study of JAGN1-dependent ER membrane integrity and stress responses, offering insights into neutropenia-associated ER defects.
Research applications include molecular characterization of ER stress pathways, pharmacological screening of UPR modulators, and functional complementation studies with wild-type or mutant JAGN1. Typical assays involve western blotting for UPR markers such as CHOP, BIP, and phosphorylated PERK; RT-qPCR monitoring of XBP1 splicing; immunofluorescence to visualize ER structural changes; and flow cytometry for quantifying apoptosis. The cells can also be used in drug sensitivity assays with ER stress inducers like tunicamycin or thapsigargin. For further technical information or to discuss your application, please contact Ascent Research.
