The ANLN Knockout HEK293T Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population designed for loss-of-function studies of the ANLN gene in a human embryonic kidney epithelial background. This polyclonal pool comprises a heterogeneous mixture of edited cells harboring disruptions in the ANLN locus, providing a robust model to investigate gene function without clonal selection biases. The knockout model enables dissection of ANLN-dependent processes in a versatile and widely used host cell line.
The HEK293T cell line originates from human embryonic kidney cells transformed with the SV40 large T antigen, which confers high transfection efficiency and robust protein expression capabilities. These epithelial cells are extensively utilized for transient and stable protein production, lentivirus and retrovirus packaging, and signaling pathway analysis. The HEK293T background supports rapid proliferation and is amenable to a broad range of genetic manipulations, making it an ideal chassis for studying cell cycle regulation and cytoskeletal dynamics.
ANLN encodes anillin, a scaffold protein that orchestrates cytokinesis by linking the plasma membrane to the contractile ring. ANLN is activated downstream of cell cycle kinases CDK1 and PLK1, and is recruited to the cleavage furrow by RhoA and its guanine nucleotide exchange factor ECT2. At the division plane, ANLN interacts with filamentous actin (ACTB), non-muscle myosin II (MYH9), and septins (SEPT2, SEPT7) to bundle F-actin and anchor the actomyosin network, driving membrane ingression. This coordinated assembly ensures proper furrow positioning and progression, with ANLN serving as a hub for RhoA- and RAC1-mediated signals. Loss of ANLN disrupts contractile ring integrity, leading to cytokinesis failure and multinucleation.
In HEK293T cells, ANLN knockout recapitulates cytokinesis defects observed in cancer and developmental disorders, providing a platform to study mechanisms underlying genomic instability and abnormal proliferation. The polyclonal nature of this population mimics heterogeneous tumor environments, enabling investigation of ANLN-dependent pathways without clonal bias. Since HEK293T cells are non-transformed yet highly proliferative, this model is particularly suited for dissecting the interplay between ANLN, cell cycle machinery, and actin regulation in a controlled epithelial context.
These polyclonal knockout cells are validated for a variety of experimental applications, including western blotting to confirm ANLN depletion, immunofluorescence imaging of cleavage furrow markers, and flow cytometry-based cell cycle analysis to quantify multi-nucleation and ploidy changes. The model is ideal for siRNA knockdown rescue experiments to assess functional domains of ANLN and for co-immunoprecipitation studies to probe protein?Cprotein interactions within the contractile ring. Researchers can employ this system in drug screening campaigns targeting cytokinesis or in mechanistic studies of RhoA?CANLN?CF-actin signaling. For further details or to inquire about custom services, please contact Ascent Research.
