The L3MBTL3 Knockout HEK293T Polyclonal Cells are a CRISPR/Cas9-edited polyclonal cell population derived from HEK293T cells, designed to disrupt endogenous L3MBTL3 expression. This heterogeneous knockout pool, comprising cells with diverse editing events, ensures a spectrum of mutations and avoids biases from clonal selection, providing a robust loss-of-function model for functional studies.
HEK293T is an immortalized human embryonic kidney epithelial cell line that stably expresses the SV40 large T-antigen, enabling episomal replication of plasmids containing the SV40 origin. As a derivative of HEK293, this line supports high-level ectopic protein expression and efficient viral production, making it a versatile host for investigating chromatin biology, transcriptional regulation, and cell cycle control.
L3MBTL3 encodes a Polycomb group protein that reads methylated histones, primarily H4K20me1 and H4K20me2, via its MBT domains. It mediates chromatin compaction and transcriptional repression of E2F-responsive cell cycle regulators and lineage differentiation genes. L3MBTL3 interacts with RB1, E2F transcription factors, and components of PRC2 (e.g., EZH2, SUZ12) as well as HDAC1/2, consolidating silencing networks. Its activity is regulated by E2F/RB signaling and post-translational modifications. Disruption of L3MBTL3 thus derepresses target genes and alters chromatin architecture.
In HEK293T cells, L3MBTL3 knockout provides a tractable system to dissect its chromatin-regulatory and tumor-suppressive roles. The heterogeneous knockout pool minimizes clonal artifacts and better captures population-level behavior, making it particularly valuable for screening epigenetic modulators or dissecting L3MBTL3??s role in cell cycle regulation and tumor suppression. The cell line??s high transfectability and protein expression capacity facilitate studies on hematologic malignancies such as AML and MDS.
This knockout product supports validation by Western blotting and RT-qPCR, chromatin accessibility profiling via ATAC-seq or MNase-seq, and co-immunoprecipitation of interacting factors such as RB1 or EZH2. Functional assays include flow cytometric cell cycle analysis and proliferation measurements (MTT, colony formation). Additionally, the cells are suited for high-throughput epigenetic drug screens and synthetic lethal interaction studies. For further information, contact Ascent Research.
