CD64 CRISPR Activation Plasmid (m): sc-420305-ACT

Mouse Fcgr1 encodes CD64 (FcγRI), a high-affinity receptor for IgG that is predominantly expressed on monocytes, macrophages, and dendritic cells, where it couples immune complex recognition to phagocytosis, oxidative burst, and cytokine production. CD64 signals through the FcR γ-chain ITAM motif to engage SYK, PI3K, MAPK, and NF-κB pathways, integrating innate immune activation with antigen uptake and processing. As a regulator of antibody-dependent effector functions, Fcgr1 is widely studied in inflammatory and autoimmune settings, infection models, and tumor-associated myeloid biology, where shifts in CD64 expression can reflect altered activation states and myeloid differentiation. Its role in immune complex handling also links Fcgr1 to complement crosstalk, Fc-mediated endocytosis, and downstream transcriptional programs that shape tissue inflammation.

CD64 CRISPR Activation Plasmid (m) provides a targeted, non-destructive approach to upregulating endogenous Fcgr1 expression without altering the underlying DNA sequence.

CD64 CRISPR Activation Plasmid (m) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the Fcgr1 locus in human cell lines. The system is built around a catalytically inactive Cas9 (dCas9) carrying two inactivating mutations (D10A and N863A) that eliminate nuclease activity while preserving DNA binding. This dCas9 is fused to VP64, a potent transcriptional activator, and is co-expressed with a blasticidin resistance gene for selection. The second plasmid encodes the MS2-p65-HSF1 fusion protein, a secondary activator complex that works in concert with dCas9-VP64, alongside a hygromycin resistance gene. The third plasmid encodes a target-specific 20 nt sgRNA fused to two MS2 RNA aptamers that recruit the MS2-p65-HSF1 complex to the activation site, accompanied by a puromycin resistance gene. The three plasmids are delivered at a 1:1:1 mass ratio for balanced expression of all system components.

Once assembled at the target locus, the SAM complex binds within approximately 200 bp upstream of the Fcgr1 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous CD64 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native Fcgr1 locus and enabling the study of CD64-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of CD64 pathway restoration in tumor cells with silenced or reduced Fcgr1 expression.

For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.