The INO80D Knockout HT29 Polyclonal Cells represent a CRISPR/Cas9-edited polyclonal knockout cell population designed for functional studies of INO80D in a human colorectal adenocarcinoma background. This product provides a loss-of-function model achieved by targeted disruption of the INO80D gene, enabling investigation of its roles in chromatin remodeling, DNA repair, and transcription regulation. The polyclonal format reflects a heterogeneous population of cells with diverse editing events, making it suitable for pooled assays and high-throughput screens that capture population-level phenotypes without clonal biases. This model does not imply monoclonality or biallelic knockout, offering a flexible tool for researchers examining INO80D-dependent processes.
The host cell line, HT29, is a well-characterized human colorectal adenocarcinoma line with epithelial morphology. Derived from a primary colorectal tumor, HT29 cells are widely used as an intestinal epithelial model for colorectal cancer research. They harbor mutations in key oncogenes and tumor suppressors, including APC, TP53, and KRAS, making them representative of the genetic landscape of colorectal tumors. Their robust growth and compatibility with diverse experimental platforms, such as 2D monolayers and 3D spheroids, render them a versatile system for studying cancer biology, drug responses, and genomic instability in the context of the intestinal epithelium.
INO80D encodes a critical subunit of the INO80 chromatin remodeling complex, which orchestrates nucleosome positioning and histone variant exchange. Mechanistically, INO80D is activated by ATM and ATR kinases in response to DNA double-strand breaks (DSBs), and it functions downstream of E2F transcription factors during cell cycle progression. The INO80 complex, including subunits such as INO80, ACTR5, and ACTR8, interacts directly with BRCA1 and RAD51, facilitating chromatin relaxation and promoting RAD51 filament formation at damage sites. This process is essential for homologous recombination repair, as INO80D recruits BRCA1 to foci and regulates ??H2AX dynamics and H2A.Z deposition at DSBs, thereby modulating chromatin accessibility and the assembly of repair complexes involving 53BP1 and MDC1.
In the HT29 colorectal adenocarcinoma model, disruption of INO80D abrogates a key chromatin remodeling activity that maintains genomic stability. Given that colorectal cancer frequently exhibits DNA repair defects and heightened replication stress, loss of INO80D sensitizes these cells to DNA-damaging agents and PARP inhibitors, providing a platform to study synthetic lethality and therapeutic vulnerabilities. This model enables dissection of how INO80D influences tumorigenic properties such as proliferation, apoptosis, and genomic instability, illuminating its potential as a target in colorectal cancer and other malignancies driven by replication stress.
Researchers can employ this polyclonal knockout population in a wide range of assays to probe DNA damage response and chromatin dynamics. Clonogenic survival assays and drug sensitivity screens with cisplatin or PARP inhibitors assess chemosensitivity, while immunofluorescence detection of ??H2AX and RAD51 foci monitors DSB repair efficiency. Western blotting quantifies INO80D, ??H2AX, and RAD51 levels, and the comet assay evaluates global DNA damage. Chromatin studies using ChIP-seq for histone modifications and H2A.Z localization, along with RNA-seq for transcriptional profiling, reveal the epigenomic and transcriptomic consequences of INO80D loss. For further information, please contact Ascent Research.
