The DTX3L Knockout K-562 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population targeting the DTX3L gene in the K-562 human myelogenous leukemia cell line. This heterogeneous pool harbors diverse gene-disrupting mutations, offering a robust loss-of-function model free from monoclonal artifacts, suitable for population-level studies of DNA damage and innate immune pathways.
K-562 is a pluripotent hematopoietic cell line derived from a 53-year-old female with chronic myelogenous leukemia in blast crisis. It retains the ability to differentiate into multiple lineages under appropriate stimuli. Characterized by the Philadelphia chromosome and high proliferative rate, K-562 is a standard model for leukemia biology, drug sensitivity assays, and signal transduction studies, particularly for exploring oncogenic signaling and tumor suppressor pathways in hematopoietic contexts.
DTX3L encodes an E3 ubiquitin ligase that, in complex with PARP9, catalyzes ubiquitination of substrates such as histone H2B to regulate DNA damage repair and interferon (IFN)-stimulated gene expression. Its expression is induced by type I and type II IFNs via JAK-STAT signaling: IFNAR1/2 receptor activation triggers JAK1/TYK2 phosphorylation of STAT1 and STAT2, which together with IRF9 form the ISGF3 complex to transactivate DTX3L. DTX3L subsequently enhances ISG expression through ubiquitination-dependent modulation of STAT1, IRF7, and NF-??B. Upon DNA damage, ATM/ATR kinases promote DTX3L localization to breaks, where it interacts with PARP1, BRCA1, and 53BP1 to facilitate repair. DTX3L also partners with UBE2D family E2 enzymes and the ligase DTX1.
In the K-562 leukemia background, DTX3L disruption enables dissection of ubiquitin-dependent signaling in leukemogenesis and drug response. As DTX3L is often overexpressed in lymphomas and leukemias, loss of function may sensitize cells to chemotherapeutic DNA-damaging agents. This model allows assessment of DTX3L??s role in genome stability, apoptosis, and interferon-driven immune evasion, providing a relevant system to study cancer cell adaptation to genotoxic stress and innate immune activation.
Key applications include monitoring DNA damage foci by ??-H2AX immunofluorescence, quantifying ISG induction via RT-qPCR or RNA-seq after IFN treatment, and co-immunoprecipitation of DTX3L-PARP9 complexes. These polyclonal cells are suitable for viability assays with drugs like cytarabine or doxorubicin, flow cytometry for apoptosis and cell cycle, and screening of ubiquitination substrates. They also support reporter assays for interferon-responsive promoters. For more information, please contact Ascent Research.
