The KAT7 Knockout NCI-H1299 Polyclonal Cells represent a CRISPR/Cas9-edited polyclonal knockout cell population designed for comprehensive loss-of-function investigation of the KAT7 gene. This product comprises a heterogeneous pool of NCI-H1299 cells carrying targeted gene disruptions, enabling functional studies without the need for single-cell cloning. The polyclonal format preserves genetic diversity while providing a robust model for interrogating KAT7-dependent processes in DNA replication, chromatin modification, and cell cycle regulation.
The parental NCI-H1299 cell line is an extensively characterized non-small cell lung carcinoma (NSCLC) model derived from a lymph node metastasis of a lung adenocarcinoma. These cells are p53-deficient and harbor wild-type KRAS, rendering them particularly valuable for studying p53-independent oncogenic mechanisms. NCI-H1299 is widely used to investigate metastatic progression, cell cycle control, and epigenetic drivers of tumorigenesis, with its p53-null status offering a sensitive background for assessing replication stress responses.
KAT7, also known as HBO1, encodes the catalytic subunit of the HBO1 histone acetyltransferase complex, which includes the scaffolding proteins BRPF1, JADE1, ING5, and EAF6. This complex specifically acetylates histone H4 at lysines 5, 8, and 12 (H4K5ac, H4K8ac, H4K12ac) at DNA replication origins, a modification essential for recruiting the origin recognition complex (ORC1-6), CDC6, and CDT1. These factors orchestrate the loading of the MCM2-7 helicase complex to license origins for replication initiation. KAT7 activity is positively regulated by E2F transcription factors and cyclin E/CDK2 kinase, linking cell cycle progression to origin firing. Disruption of KAT7 therefore impairs MCM loading and DNA replication, culminating in stalled S-phase entry and proliferation defects.
In the NCI-H1299 p53-null context, knockout of KAT7 is particularly impactful because these cells lack p53-dependent checkpoints that normally mitigate replication stress. Loss of KAT7-driven origin licensing is predicted to exacerbate replication fork stalling, reduce cell proliferation, and increase genomic instability. This model thus provides a powerful platform for dissecting the interplay between histone acetylation and cell cycle control in a clinically relevant lung cancer background. Given the prevalence of p53 deficiency in NSCLC, this system is well-suited for exploring KAT7 as a candidate therapeutic target in tumors reliant on robust replication licensing.
Research applications include functional analysis of DNA replication licensing, epigenetic regulation of G1/S transition, and target validation in p53-null cancers. Representative assays facilitated by this model are western blotting for histone H4 acetylation markers (H4K5ac, H4K8ac, H4K12ac), RT-qPCR for KAT7 transcript, cell viability (MTS) and proliferation (EdU) assays, flow cytometry with PI staining for cell cycle, DNA fiber assays, co-immunoprecipitation of the HBO1 complex, and ChIP-qPCR to measure H4K5ac at origins. For further technical details, contact Ascent Research.
