The HASPIN Knockout HCT 116 Polyclonal Cells provide a robust loss-of-function model generated by CRISPR/Cas9-mediated disruption of the HASPIN gene in HCT 116 colorectal carcinoma cells. As a polyclonal knockout cell population, this product consists of a heterogeneous pool of edited cells, each carrying targeted gene disruption, thereby avoiding the artifacts associated with single-cell cloning while preserving the biological variability inherent in tumor populations. This format is particularly suitable for functional genomics studies, drug screening, and phenotypic analyses where maintaining population-level responses is critical.
The host cell line, HCT 116, is a well-characterized human colorectal carcinoma epithelial model with microsatellite instability (MSI), a KRAS G13D mutation, and a CTNNB1 (??-catenin) mutation, resulting in mismatch repair deficiency. These genetic features make HCT 116 cells highly relevant for studying colorectal adenocarcinoma pathogenesis, including Wnt signaling dysregulation and chromosomal instability phenotypes. The parental line is widely used in cancer research for its rapid proliferation and reproducible responses to chemotherapeutic agents.
HASPIN is a serine/threonine kinase that operates as a critical mitotic checkpoint regulator. The enzyme is activated by the CDK1/cyclin B complex and phosphorylates histone H3 at threonine 3 (H3T3) at inner centromeres. This phosphorylation event creates a docking site for the chromosomal passenger complex (CPC) through direct interaction with Survivin (BIRC5), which consequently recruits Aurora B kinase to centromeric regions. HASPIN??s activity is further modulated by PLK1 and it functions upstream of Aurora B, INCENP, and Borealin, forming a signaling axis that coordinates centromeric cohesion protection and proper chromosome alignment. Disruption of HASPIN abolishes H3T3 phosphorylation and mislocalizes the CPC, leading to severe mitotic defects.
In the HCT 116 background, the HASPIN knockout is particularly informative because the host cell??s intrinsic MSI and defective mismatch repair create a sensitized environment for chromosomal instability. The loss of HASPIN??s kinase activity exacerbates errors in chromosome segregation and mitotic progression, potentially unmasking synthetic lethal interactions or enhancing sensitivity to agents targeting the spindle assembly checkpoint. This model allows researchers to dissect how oncogenic mutations cooperate with mitotic dysregulation in colorectal cancer and may reveal therapeutic vulnerabilities specific to tumors with centrosome amplification or cohesion defects.
This polyclonal knockout product is designed for diverse research applications, including mitotic checkpoint disruption studies, chromosomal instability assays, and kinase inhibitor screening. Representative experimental approaches include western blotting for H3T3ph to confirm target engagement, immunofluorescence microscopy to assess Aurora B localization and centromere dynamics, flow cytometry for cell cycle profiling, colony formation assays to evaluate proliferation, and TUNEL assays to measure apoptosis. The model is also well-suited for synthetic lethality screens and evaluating compounds that target the HASPIN?CCPC signaling node. Researchers are encouraged to contact Ascent Research for further technical details and custom applications.
