The GFAP Knockout HEK293T Polyclonal Cells are a CRISPR/Cas9-edited polyclonal population with targeted disruption of the GFAP gene. This heterogeneous knockout model enables robust functional studies without clonal bias, providing a practical tool for investigating GFAP-mediated processes in a widely utilized human cell background.
HEK293T cells are human embryonic kidney epithelial cells immortalized with SV40 large T antigen, offering high transfection efficiency, rapid proliferation, and reliable protein expression. Derived from HEK293 cells, this line is a cornerstone of biomedical research, especially for viral production and protein characterization, ensuring reproducibility across experiments. Its well-characterized biology and straightforward culture conditions make it an ideal platform for CRISPR-based gene disruption.
GFAP encodes glial fibrillary acidic protein, a type III intermediate filament constituting the astrocyte cytoskeleton. It assembles with vimentin, nestin, and synemin into networks anchored by plectin and integrins. GFAP expression is induced by STAT3 downstream of JAK2-activated cytokine receptors (e.g., LIF, CNTF), by Notch1/RBP-J signaling, and by BMP-triggered Smad1/5/8. NF-??B also contributes. In turn, GFAP influences expression of vimentin, tenascin-C, and chondroitin sulfate proteoglycans, which drive reactive gliosis. Thus, GFAP knockout eliminates intermediate filaments and uncouples these signaling pathways from cytoskeletal outputs.
Within the HEK293T background, GFAP loss disrupts intermediate filament networks that normally provide mechanical support and signaling integration. This model allows dissection of GFAP??s roles in cytoskeletal dynamics and its crosstalk with JAK-STAT, Notch, and BMP pathways outside the astrocytic context. The polyclonal nature captures diverse mutations, ensuring that observed phenotypes are consistent and not artifacts of clonal selection.
Applications include functional characterization of GFAP mutations implicated in Alexander disease, antibody validation by Western blotting and immunofluorescence, drug sensitivity screening targeting intermediate filaments, and cell migration assays. It also supports intermediate filament assembly studies, as well as genotype and expression analysis via Sanger sequencing and RT-qPCR. Additionally, it serves as a simplified system to study reactive gliosis mechanisms. For further information, contact Ascent Research.
