The KLF4 Knockout HEK293T Polyclonal Cells represent a CRISPR/Cas9-edited polyclonal knockout cell population derived from the HEK293T human embryonic kidney cell line, featuring targeted disruption of the KLF4 gene. This pool of knockout cells provides a heterogeneous loss-of-function model suitable for studying the functional roles of KLF4 in a robust cellular context. Unlike clonal isolates, the polyclonal format maintains genetic diversity, enabling comprehensive analysis of KLF4-dependent phenotypes while minimizing clonal artifacts. The cells are delivered as a ready-to-use population, supporting immediate application in downstream assays relevant to cancer biology, stem cell research, and signal transduction studies.
The host cell line, HEK293T, is a well-characterized derivative of human embryonic kidney epithelial cells that stably expresses the SV40 large T antigen. This modification permits high-level episomal replication of plasmids containing the SV40 origin of replication, making the line exceptionally transfectable and widely used for exogenous protein expression, lentiviral production, and functional genomics. The epithelial origin and robust growth characteristics of HEK293T cells provide a physiologically relevant and experimentally tractable backdrop for investigating gene function, particularly for transcription factors like KLF4 that govern epithelial cell identity and proliferation.
KLF4 (Kr??ppel-like factor 4) is a zinc-finger transcription factor that binds GC-rich DNA elements to regulate genes critical for cell cycle progression, differentiation, pluripotency, and epithelial barrier integrity. It exhibits a context-dependent dual role in cancer, functioning as either a tumor suppressor or an oncogene. KLF4 is activated by upstream regulators including p53, SP1, STAT3, and ??-catenin/TCF, and it transcriptionally modulates key downstream targets such as CDKN1A (p21), CCND1 (cyclin D1), BAX, and CDH1 (E-cadherin). It interacts with cofactors p300/CBP and HDAC1, integrating signals from the TGF-??, WNT, MAPK/ERK, and p53 pathways. Mechanistically, KLF4 promotes p21 expression downstream of p53 to suppress proliferation, or facilitates G1/S transition via cyclin D1 regulation.
The HEK293T background is significant for KLF4 study, as SV40 large T antigen inactivates p53 and Rb, sensitizing the system to KLF4-dependent proliferation and apoptosis. The epithelial context permits analysis of KLF4??s role in barrier integrity and EMT, given its regulation of E-cadherin. High transfectability enables complementation experiments and luciferase reporter assays for transcriptional activity.
This KLF4 knockout cell pool enables studies of transcriptional regulation of proliferation and differentiation, pluripotency induction, and tumor suppression/oncogenesis. Assays include western blotting, RT-qPCR, Sanger sequencing, proliferation, apoptosis, cell cycle, luciferase reporter, and migration/invasion assays. These cells are suitable for screening KLF4 modulators. For further information, contact Ascent Research.
