GPATCH2L Knockout Jurkat Polyclonal Cells are a CRISPR/Cas9-edited polyclonal population derived from Jurkat T lymphocytes, with disrupted GPATCH2L gene function. This polyclonal knockout model enables loss-of-function studies of GPATCH2L in DNA damage signaling and repair. CRISPR/Cas9-mediated gene disruption creates a heterogeneous cell pool lacking full-length GPATCH2L, avoiding clone-specific artifacts and preserving varied editing outcomes for robust analysis.
The Jurkat E6.1 line originates from an acute T cell leukemia patient and is a classic model for T cell receptor signaling, cytokine production, and apoptosis. Its p53-null status and responsive TCR pathways make it valuable for dissecting DNA damage responses independent of p53-mediated apoptosis, emphasizing alternative repair factors like GPATCH2L.
GPATCH2L encodes a G-patch domain protein critical for non-homologous end joining (NHEJ) repair of DNA double-strand breaks (DSBs). It is activated by ATM, ATR, and DNA-PKcs upon damage and directly interacts with Ku70/Ku80 and DNA-PKcs at break sites. This scaffolding facilitates recruitment of XRCC4, LIG4, and 53BP1, while promoting H2AX phosphorylation (??H2AX). GPATCH2L is regulated by CDK1 and p53; its depletion results in persistent ??H2AX foci, CHK1/CHK2 checkpoint activation, and increased sensitivity to genotoxic stress.
In Jurkat cells, GPATCH2L disruption unveils NHEJ defects without p53-mediated apoptosis interference. Polyclonal knockout cells accumulate ??H2AX and 53BP1 foci after irradiation or radiomimetic treatment, confirming impaired DSB repair. This phenotype enables exploration of genetic interactions with ATM, DNA-PKcs, or PARP1 inhibitors. The model is highly relevant for studying DNA repair-dependent survival, clonal evolution, and therapeutic resistance in T cell leukemia and genomic instability syndromes.
These cells support numerous assays: Western blot for ??H2AX and phospho-ATM; immunofluorescence visualization of 53BP1 foci; comet and colony formation assays for genotoxic sensitivity; flow cytometry for cell cycle and apoptosis; co-immunoprecipitation to assess GPATCH2L?CKu70/DNA-PKcs interactions; and RT-qPCR or RNA-seq for transcriptional profiling of p21, BAX. Applications include DNA damage repair mechanistics, T cell leukemia functional genomics, DNA repair inhibitor screening, cancer drug resistance research, and biomarker validation. For further information, contact Ascent Research.
