The HAT1 Knockout HT29 Polyclonal Cells provide a CRISPR/Cas9-mediated gene-disrupted polyclonal population targeting the HAT1 locus in the HT29 human colorectal adenocarcinoma cell line. This heterogeneous pool of edited cells serves as a loss-of-function model, enabling investigation of HAT1-dependent processes without selection for a single clonal genotype. The knockout is introduced across a bulk cell population, maintaining polyclonal representation that can be employed in pooled functional genomic and epigenetic studies to assess average phenotypic outcomes.
HT29 cells are a widely utilized epithelial colorectal adenocarcinoma model established from a female patient. These adherent cells display epithelial morphology and are characterized by mutations in key oncogenic and tumor-suppressor pathways, including APC and KRAS alterations, making them a standard platform for colorectal cancer research. Their capacity to form tumors in xenograft assays and distinct differentiation properties under post-confluent conditions further extend their utility in studying intestinal tumor biology, metastasis, and therapeutic response.
HAT1 encodes a type B histone acetyltransferase that acetylates newly synthesized histone H4 on lysine 5 (H4K5ac) and lysine 12 (H4K12ac), a modification critical for chromatin assembly during DNA replication and repair. HAT1 activity is tightly coupled to the cell cycle and DNA replication machinery: it is transcriptionally activated by E2F1 and MYC, and functionally linked to proliferating cell nuclear antigen (PCNA). Within the chromatin assembly pathway, HAT1 interacts with the histone chaperone complex p46/p48 (RbAp46/RbAp48), chromatin assembly factor 1 (CAF-1), and Asf1 to deposit acetylated H3?CH4 dimers onto nascent DNA. This acetylation mark facilitates proper chromatin maturation and genome stability, and its dysregulation can lead to aberrant gene expression patterns and oncogenic transformation.
In the context of HT29 colorectal cancer cells, HAT1 disruption offers a direct means to dissect replication-coupled chromatin assembly and its consequences on the epigenetic landscape of an intestinal tumor model. Given the reported overexpression of HAT1 in colorectal, lung, and hepatocellular carcinomas, this knockout model enables functional interrogation of its putative tumorigenic role. Loss of HAT1 is expected to reduce H4K5ac and H4K12ac marks, impair proper chromatin maturation, and may perturb expression of genes governing proliferation, adhesion, and drug sensitivity, providing mechanistic insight into HAT1-driven oncogenic processes.
Researchers can apply this knockout model to a range of experimental paradigms, including Western blotting for HAT1 and histone acetylation marks, chromatin immunoprecipitation?CqPCR (ChIP?CqPCR) for H4K5ac and H4K12ac occupancy, immunofluorescence to monitor chromatin markers, and cell cycle analysis by flow cytometry. Functional assays such as migration/invasion tests and HDAC inhibitor sensitivity screens complement transcriptomic profiling via RNA-seq. For additional information, please contact Ascent Research.
