The HLTF Knockout Jurkat Polyclonal Cells comprise a CRISPR/Cas9-edited polyclonal knockout population harboring disrupted HLTF alleles. This heterogeneous pool provides a versatile loss-of-function model for studying HLTF biology without clonal artifacts. The polyclonal nature captures a range of editing outcomes, enabling robust functional screens and comparative analyses in the context of human T-lymphocyte biology.
Jurkat is an immortalized T-cell line isolated from an acute T-cell leukemia patient. Widely adopted for studying T-cell signaling, apoptosis, and oncogenic transformation, these cells possess functional DNA damage response pathways, including ATR/Chk1 and homologous recombination machineries. Their transformed background mimics aspects of tumor biology, making them particularly relevant for investigating replication stress responses and genomic instability mechanisms.
HLTF functions as a DNA translocase and ubiquitin ligase that mediates replication fork reversal and DNA damage tolerance, thereby preventing genomic instability and suppressing tumorigenesis. Catalytic activity is stimulated by ATR-mediated phosphorylation and regulated by p53 and E2F transcriptional control. HLTF directly binds PCNA and RAD18 at stalled forks, promotes fork remodeling, and influences expression of DNA repair genes such as RAD51. Its loss impairs damage resolution, leading to elevated double-strand breaks and heightened sensitivity to genotoxins.
In Jurkat cells, HLTF disruption enables dissection of how T-cell replication stress intersects with lymphomagenesis and treatment resistance. As HLTF is mutated or silenced in colorectal, esophageal, and breast cancers, this model facilitates mechanistic studies of HLTF tumor-suppressive roles and may reveal synthetic lethal partners in DNA repair-deficient backgrounds. The cells are valuable for testing drug sensitivities and evaluating the interplay between oncogenic stress and DNA damage checkpoints.
These polyclonal cells are suited for numerous assays, including western blotting for HLTF and downstream markers (??H2AX, RAD51), immunofluorescence for DNA damage foci, comet assays, and viability/clonogenic survival assays following genotoxic challenge. Flow cytometry enables cell cycle analysis; co-immunoprecipitation maps interactions with PCNA and RAD18; ChIP-qPCR examines chromatin occupancy. Applications span DNA repair, cancer biology, and synthetic lethality screening. For additional information, please contact Ascent Research.
