The ITPR1 Knockout Jurkat Polyclonal Cells consist of a CRISPR/Cas9-edited polyclonal population derived from the Jurkat T-lymphocyte cell line, with targeted disruption of the ITPR1 gene. This bulk knockout model preserves the inherent heterogeneity of the parental line while providing a robust loss-of-function system to investigate ITPR1-dependent calcium signaling, avoiding clonal selection artifacts.
Jurkat cells are a human T-cell acute lymphoblastic leukemia (T-ALL) line widely used for studying T-cell receptor (TCR) signal transduction, activation-induced calcium flux, and apoptosis. Their genetic malleability and well-defined signaling pathways make them an optimal host for CRISPR-mediated gene disruption, enabling detailed dissection of calcium-regulated T-cell functions.
ITPR1 encodes the IP3 receptor type 1, an endoplasmic reticulum calcium channel that mediates intracellular Ca2+ release in response to IP3. In T cells, TCR ligation activates PLC??1, which hydrolyzes PIP2 to produce IP3. IP3 binding to ITPR1 triggers Ca2+ efflux, activating calmodulin and calcineurin, which then dephosphorylates NFAT transcription factors, promoting their nuclear entry and transcriptional activity. ITPR1 function is modulated by interacting proteins including IRBIT, FKBP12, and HOMER, and it connects to downstream effectors such as CaMKII, calpain, and the mitochondrial calcium uniporter.
In Jurkat cells, ITPR1 knockout ablates the principal IP3-dependent calcium release mechanism, making it an ideal model to dissect calcium??s role in T-cell activation, proliferation, and apoptosis. This system also aids in exploring calcium-dependent oncogenic pathways in T-ALL and can serve as a platform to study ITPR1-related signaling defects linked to spinocerebellar ataxia type 15/16 and Gillespie syndrome.
Typical applications include Fluo-4?Cbased calcium flux measurements, flow cytometry for activation markers (CD69, CD25), Western blotting for phospho-PLC??1 and calcineurin activity, and NFAT luciferase reporter assays. Additionally, annexin V apoptosis and CFSE proliferation assays enable comprehensive functional analysis. For further details or technical inquiries, please contact Ascent Research.
