Innate immunity functions as the first line against infection, which is mediated by series of innate immune receptors. RIG-I like receptors (RLR) recognize the cytosolic nucleic acids mainly from viruses, trigger the type I interferon (IFN) production, and thus play essential roles in the anti-viral immunity. Here we cloned the mouse RIG-I (mRIG-I) cDNA into the pENTR vector using the TOPO cloning technology, and transferred the FLAG tagged mRIG-I gene from pENTR vector to destination pLenti viral vector. The expression of mRIG-I in transfected cells was detected by immunoblotting using anti-FLAG and anti-RIG-I antibodies. The transfected mRIG-I was active capable of inducing downstream IFN stimulated gene (ISG) gene transcription as well as protein expression.
Keywords: Gateway cloning; ISG; mouse RIG-I; TOPO cloning
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