CD16 CRISPR Activation Plasmid (h): sc-400544-ACT

FCGR3A encodes CD16 (FcγRIIIa), a low-to-intermediate affinity receptor for IgG Fc that is prominently expressed on natural killer cells and subsets of monocytes/macrophages. CD16 couples to ITAM-bearing adaptor chains (FCER1G and CD247) to initiate signaling cascades involving SYK, PI3K/AKT, PLCγ, and MAPK, integrating antibody-dependent cellular cytotoxicity, cytokine production, and immune complex handling. This receptor participates in innate immune activation, inflammatory regulation, and crosstalk with interferon and NF-κB programs that shape effector cell function. Altered FCGR3A activity and cell-surface expression have been associated with dysregulated immune responses in autoimmunity, chronic inflammation, infection, and tumor immunology, supporting mechanistic studies in human immune cell models.

CD16 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous FCGR3A expression without altering the underlying DNA sequence.

CD16 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the FCGR3A 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 FCGR3A transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous CD16 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native FCGR3A locus and enabling the study of CD16-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of CD16 pathway restoration in tumor cells with silenced or reduced FCGR3A expression.

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