The EHMT2 Knockout HCT 116 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population generated from the HCT 116 human colorectal adenocarcinoma cell line. This product provides a mixed population of cells harboring targeted disruption of the EHMT2 gene, enabling loss-of-function studies without the selection of a single clonal isolate. The polyclonal format reflects the heterogeneity of knockout events within the cell pool, making it well-suited for bulk functional assays where population-level effects are of interest.
The host cell line, HCT 116, is a well-characterized epithelial model of colorectal carcinoma. It exhibits high microsatellite instability (MSI-H) and carries a KRAS G13D oncogenic mutation, features that are prevalent in a subset of colorectal tumors. The cell line has a near-diploid karyotype and is extensively used to study mechanisms of tumorigenesis, cancer cell signaling, and responsiveness to chemotherapeutic and targeted agents.
EHMT2 (also known as G9a) encodes a histone lysine methyltransferase responsible for catalyzing mono- and dimethylation of histone H3 at lysine 9 (H3K9me1/2), a modification associated with transcriptional silencing and heterochromatin formation. EHMT2 activity is regulated by upstream factors including E2F1, MYC, and p53, and it mediates the epigenetic repression of key tumor suppressor genes such as CDH1 (E-cadherin), DLC1, RUNX3, PTEN, and CDKN1A (p21). Functionally, EHMT2 interacts with diverse partners including DNMT1, HP1??, UHRF1, CBX5, EZH2, HDAC1/2, GLP/EHMT1, LSD1, and transcriptional repressors SNAIL, ZEB1, and CTBP to reinforce gene silencing. In the context of the Wnt/??-catenin pathway, EHMT2 cooperates with ??-catenin/TCF4 complexes, while also modulating TGF-??/SMAD and p53/p21/MDM2 signaling axes.
In HCT 116 cells, EHMT2 overexpression contributes to the epigenetic suppression of epithelial markers and the activation of pro-invasive transcriptional programs. Loss of EHMT2 function in this MSI-H, KRAS-mutant background is predicted to relieve repression of CDH1 and PTEN, thereby potentially reversing aspects of epithelial-mesenchymal transition and dampening tumor cell proliferation and survival. The HCT 116 line??s intact yet dysregulated Wnt/??-catenin pathway makes it particularly relevant for examining how EHMT2 influences ??-catenin?Cdependent transcription and its downstream effects on cell adhesion and motility.
The EHMT2 Knockout HCT 116 Polyclonal Cells support a broad range of research applications. They can be employed in functional studies of EHMT2-mediated gene silencing using chromatin immunoprecipitation (ChIP-qPCR) to assess H3K9me2 enrichment at target gene promoters, combined with RT-qPCR or RNA-seq transcriptomic profiling to measure derepression of genes like CDH1, PTEN, and CDKN1A. Western blotting for H3K9me1/2 provides direct readouts of catalytic activity loss. The cells are suitable for investigating the role of EHMT2 in colorectal cancer EMT and metastasis through wound healing or transwell migration/invasion assays, and for studying DNA damage response and autophagy regulation. Additionally, they serve as a platform for epigenetic drug screening, including sensitivity testing with EHMT2 inhibitors such as BIX-01294 or UNC0642, either alone or in combination with demethylating agents. Apoptosis (Annexin V/PI) and co-immunoprecipitation can further elucidate functional consequences and protein interaction networks. For questions regarding this product, please contact Ascent Research.
