The ARPP21 Knockout HEK293T Polyclonal Cells represent a CRISPR/Cas9-edited cell population carrying targeted disruption of the human ARPP21 gene. This polyclonal pool consists of HEK293T cells harboring heterogeneous indel mutations at the ARPP21 locus, generating a functional knockout model suitable for bulk analyses. The polyclonal format reduces clonal biases and ensures robust representation of loss-of-function genotypes, facilitating reproducible biochemical and signaling studies.
HEK293T cells are an adenovirus 5-transformed human embryonic kidney epithelial line that stably expresses the SV40 large T-antigen. This background permits high-level transient protein expression and efficient amplification of SV40 origin-containing plasmids, making it a preferred host for reconstituting signaling pathways and producing recombinant proteins or lentiviral vectors. The adherent monolayer growth and facile transfectability of HEK293T cells suit them for knockout-based functional assays.
ARPP21 is a cAMP-regulated phosphoprotein that acts as a key node in dopamine D1 receptor??PKA??PP1 signaling cascades. Upon D1 receptor activation, elevated cAMP activates PKA, which phosphorylates ARPP21 at Ser55. Phosphorylated ARPP21 inhibits protein phosphatase 1 (PP1, catalytic subunit PPP1CA), thereby modulating dephosphorylation of targets like DARPP-32 and other phosphoproteins. This regulatory loop is central to striatal medium spiny neuron plasticity, and ARPP21 is enriched in the striatum.
While HEK293T cells do not natively express dopamine receptors, they offer a clean cellular background to reconstitute ARPP21-dependent pathways via transient or stable introduction of components such as DRD1. The knockout context allows unambiguous assessment of ARPP21’s contribution to cAMP/PKA-driven PP1 inhibition without interference from endogenous protein. This makes the cells an attractive system for dissecting ARPP21-mediated phospho-regulation and testing chemical modulators of the pathway.
These polyclonal knockout cells are intended for research into dopamine-linked disorders??including Parkinson’s disease, schizophrenia, substance use disorder, and L-DOPA-induced dyskinesia??as well as basic mechanism studies of cAMP/PKA/PP1 signaling. Common downstream assays include Western blotting, RT-qPCR, Sanger sequencing for mutation validation, cAMP accumulation measurements, PKA activity assays, co-immunoprecipitation, and phosphoproteomic profiling. For further details or to place an order, please contact Ascent Research.
