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HEK STING Reporter Cells

293T-DualSTING (ISG/KI-IFNb) cells are a family of reporter cells designed to study variants of STING (stimulator of interferon genes; also known as TMEM173, MITA, MPYS, and ERIS).

STING is essential for the interferon (IFN) response to cytoplasmic foreign or self-DNA and directly senses cyclic dinucleotides (CDNs), which are important messengers in bacteria and innate immune agonists in mammals.

Interestingly, a variety of natural non-synonymous variants of human STING that affect CDN recognition and signal transduction have been identified [1].

293T-DualSTING (ISG/KI-IFNb) cells enable the study of STING variation by monitoring the activation of the transcription factor ISRE (IFN-stimulated response elements) and/or the expression of IFN‑β.

293T-Dual™ STING (ISG/KI-IFNb) cells were generated from 293T-Dual™ Null (ISG/KI-IFNb) cells, which derive from 293T cells, human embryonic kidney 293-derived cells that contain the SV40 T-antigen and do not respond to stimulation by CDNs.

293T-Dual™ STING and their parental cell line stably express an ISRE-inducible SEAP (secreted embryonic alkaline phosphatase) reporter construct. They also express Lucia luciferase, a secreted luciferase, placed under the control of the endogenous IFN-β promoter; the coding sequence of IFN-β has been replaced by the Lucia luciferase ORF using knockin technology.

Thus, CDN stimulation can be assessed in 293T-Dual™ STING (ISG/KI-IFNb) cells by monitoring ISRE-induced SEAP production and/or IFN-β-dependent expression of Lucia luciferase.

The two reporter proteins, SEAP and Lucia Luciferase, can be readily measured in the supernatant by using QUANTI-Blue™ and QUANTI-Luc™, respectively.

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