GGACT Knockout Raji Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout population derived from the Raji B-lymphoblastoid cell line, featuring targeted disruption of the GGACT gene. This polyclonal pool comprises cells with heterogeneous editing events, serving as a bulk loss-of-function model without clonal isolation. It is designed for population-level analyses of GGACT-dependent pathways.
The Raji cell line is an Epstein-Barr virus-positive Burkitt lymphoma-derived lymphoblastoid line of B lymphocyte origin, capable of antibody production and reflective of adaptive immune function. These cells are extensively used in B-cell malignancy research, including studies of oncogenic signaling, drug resistance, and viral-host interactions. Their EBV-driven background alters metabolic and transcriptional programs, providing a suitable context for examining glutathione metabolism and apoptosis regulation.
GGACT catalyzes conversion of gamma-glutamylamines to 5-oxoproline and free amines within the gamma-glutamyl cycle, directly regulating glutathione turnover. GGACT is transcriptionally regulated by NF-??B and STAT3, and functions in concert with gamma-glutamyltranspeptidase (GGT) and glutathione S-transferases (GSTs). Its product, 5-oxoproline, is hydrolyzed by 5-oxoprolinase, and the cycle feeds into glutathione biosynthesis via glutamate-cysteine ligase (GCL) and glutathione synthetase (GSS). Disruption of GGACT impairs this cycle, potentially reducing glutathione levels, elevating reactive oxygen species, and modulating apoptotic regulators downstream.
In Raji lymphoma cells, GGACT knockout is expected to disrupt gamma-glutamyl cycle homeostasis, potentially sensitizing cells to oxidative stress and altering apoptotic thresholds. Given that NF-??B and STAT3 are frequently activated in Burkitt lymphoma and directly regulate GGACT, this model enables dissection of transcription-dependent redox control. The polyclonal cell pool is valuable for probing how changes in glutathione metabolism influence proliferation and drug sensitivity in a B-lymphoblastoid setting.
Researchers can apply these cells to study glutathione metabolism and redox biology in lymphoma using Western blotting for GGACT, intracellular glutathione measurement, flow cytometric apoptosis assays (Annexin V/PI), ROS detection with DCFDA, and cell proliferation via MTT. Key applications include investigating chemoresistance mechanisms, redox-dependent signaling, and testing modulators of the gamma-glutamyl cycle. For further inquiries, please contact Ascent Research.
