CBFB Knockout HEK293T Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population targeting the human CBFB gene in HEK293T cells. The cell pool harbors variable targeted disruptions of CBFB, enabling population-level loss-of-function studies without clonal isolation. This model facilitates investigation of CBFB-dependent transcriptional regulation, protein interactions, and signal transduction in a widely used expression host.
HEK293T is an adherent human embryonic kidney line expressing SV40 large T-antigen, supporting episomal replication of SV40-origin plasmids and enabling high-efficiency transient transfection as well as lentiviral/retroviral production. Its robust growth and ease of genetic manipulation make it a favored background for studying protein overexpression, reporter assays, and complex formation, providing a clean cellular context for dissecting CBFB-mediated mechanisms.
CBFB encodes the non-DNA-binding beta subunit of the core-binding factor heterodimer. It allosterically stabilizes RUNX transcription factors??primarily RUNX1??on DNA, thereby driving expression of genes essential for definitive hematopoiesis, lymphocyte development, and bone formation. Upstream regulators include TGF-beta receptors, BMP receptors, NOTCH receptors, and WNT/Frizzled receptors. Downstream, the CBFB?CRUNX1 complex transactivates target genes such as CSF1R, LEF1, CD4, CD8A, IL3, and CSF2 (GM-CSF). Key interacting factors comprise RUNX2, RUNX3, SMADs, p300/CBP, and TLE/HDAC corepressors, contextualizing CBFB within multiple developmental signaling networks and leukemogenic pathways.
While HEK293T lacks the hematopoietic context where CBFB functions primarily, its heterologous knockout allows precise biochemical dissection of CBFB-dependent transactions. Reporter assays for RUNX1-mediated transactivation benefit from reduced background of endogenous regulators. Co-transfection experiments comparing knockout and wild-type cells clarify CBFB roles in stabilizing RUNX1?CDNA binding and cofactor recruitment. The model also permits analysis of leukemogenic CBFB?CMYH11 fusion proteins, aiding mechanistic studies of core-binding factor AML.
Applications include co-immunoprecipitation for mapping CBF?CRUNX complex assembly, ChIP-qPCR for RUNX1 locus occupancy at targets like CSF1R and LEF1, and dual luciferase reporter assays to quantify transcriptional activity changes. Combining the knockout pool with cDNA or CRISPR libraries supports synthetic lethality screens for CBF-related leukemias. Medium-throughput small-molecule screens can identify agents that modulate CBF function. For further details, custom modifications, or technical support, please contact Ascent Research.
