Rragc Knockout RAW 264.7 Cell Line

The Rragc Knockout RAW 264.7 Cell Line is a CRISPR/Cas9-edited knockout cell line generated by targeted disruption of the Rragc gene in murine RAW 264.7 macrophages. This loss-of-function model enables mechanistic studies of Rragc-dependent signaling events within a well-characterized innate immune cell context, making it valuable for functional genomics and signal transduction research.

RAW 264.7 cells are an adherent, Abelson murine leukemia virus-transformed macrophage line derived from BALB/c mice. They exhibit characteristic macrophage features, including responsiveness to lipopolysaccharide (LPS) and other pattern-associated molecular patterns, robust phagocytic activity, and antigen presentation capacity. Widely used to model innate immune responses, these cells provide a physiologically relevant host for examining how metabolic regulators like Rragc influence macrophage functions such as cytokine production, inflammasome activation, and metabolic reprogramming. Their well-defined signal transduction pathways make them ideal for dissecting the intersection of nutrient sensing and immunity.

Rragc encodes a small GTPase that forms heterodimers with RragA or RragB to regulate mTORC1 lysosomal localization. The Ragulator?Cv-ATPase complex and amino acid sensors such as Sestrin2 and CASTOR1 control Rag nucleotide loading in response to leucine and arginine. Once amino acids are abundant, the Rag heterodimer recruits mTORC1 to lysosomes, where it is activated by Rheb and subsequently phosphorylates downstream targets including S6K1, 4E-BP1, and ULK1. This signaling axis promotes protein synthesis and ribosome biogenesis while inhibiting autophagy. Growth factor signals via insulin and IGF-1 further fine-tune mTORC1 activity through parallel cascades, highlighting the integrated nature of nutrient and growth factor sensing.

In macrophages, mTORC1 activity is pivotal for translating nutrient availability into appropriate immune effector functions. Knockout of Rragc in RAW 264.7 cells is anticipated to disrupt amino acid?Cmediated mTORC1 activation, leading to defective autophagy, altered translational control, and skewed inflammatory cytokine output. This model thus offers a powerful platform to dissect the interplay between cellular metabolism and innate immunity. It also holds direct relevance for studying dysregulated mTORC1 signaling in human pathologies such as Birt-Hogg-Dub?? syndrome, where folliculin mutations impair Rag-mediated mTORC1 regulation, as well as renal cell carcinoma and follicular lymphoma, where mTORC1 hyperactivation is a common feature.

Researchers can apply this cell line to a variety of experimental approaches, including immunoblotting for phosphorylated mTORC1 targets, LC3-based autophagy flux assays, and immunofluorescence monitoring of mTORC1 lysosomal translocation. Functional studies encompass amino acid starvation and refeeding, cytokine profiling, and phagocytosis measurements. The model supports drug screening against the Rag?CmTORC1 pathway and mechanistic dissection of mutant phenotypes. For technical specifications, lot-specific genome editing verification data, or ordering information, please contact Ascent Research.