The BATF3 Knockout HT29 Polyclonal Cells product supplies a CRISPR/Cas9-edited polyclonal knockout cell population originating from the HT29 human colon adenocarcinoma cell line. This loss-of-function model is produced by disrupting the BATF3 gene via CRISPR/Cas9, yielding a heterogeneous pool of cells that avoids clonal selection biases and enables robust investigation of BATF3-dependent processes in a colorectal cancer context.
HT29 cells were established from a primary tumor of a 44-year-old Caucasian female with colon adenocarcinoma. They display epithelial morphology, mucin secretion, and the ability to form polarized monolayers, making them a standard in vitro model for colorectal cancer biology and intestinal epithelial barrier function. Their well-characterized signaling networks and responses to therapeutics render HT29 an optimal host for gene-editing studies.
BATF3 encodes a basic leucine zipper transcription factor that forms heterodimers with JUN, JUNB, or JUND to bind AP-1 consensus sequences, thereby regulating target gene transcription. Upstream regulators include IL-12, IFN-??, EGF, TGF-??, IRF8, SPI1, and WNT3A, while downstream targets encompass CCND1, BCL2, SNAI1, CDH1, MMP9, and CXCL8. BATF3 also interacts with FOS, ATF2, IRF4, and IRF8, and its activity is influenced by MAPK/ERK, JNK, Wnt/??-catenin, and TGF-?? signaling cascades. Within HT29 cells, BATF3 knockout results in reduced proliferation and increased susceptibility to apoptosis inducers, consistent with its role in promoting cell survival and cell cycle progression through AP-1-mediated transcription.
In the HT29 background, BATF3 disruption permits detailed dissection of colorectal cancer-relevant signaling networks. The knockout cells can be used to examine MAPK pathway dynamics involving MAPK8, MAP2K4, JUN, FOS, ATF2, ELK1, and DUSP1, as well as crosstalk with TGF-?? and Wnt pathways. This model is particularly suited for studying the transcriptional control of epithelial-mesenchymal transition and apoptosis resistance, as BATF3 modulates key effectors such as SNAI1, MMP9, BCL2, and CDH1.
Typical applications of this polyclonal knockout product include functional analyses of proliferation, apoptosis, and migration using MTT, Annexin V/PI, and Transwell assays. Gene expression changes can be profiled by RNA-seq and RT-qPCR, complemented by protein detection via Western blotting. Additional assays include co-immunoprecipitation for protein interactions, luciferase reporter assays to quantify AP-1 activity, and drug sensitivity screening with 5-fluorouracil or oxaliplatin. Co-culture experiments can explore BATF3’s role in tumor immune evasion. For further information or technical assistance, please contact Ascent Research.
