GCSFR CRISPR Activation Plasmid (h): sc-402269-ACT

CSF3R encodes the human granulocyte colony-stimulating factor receptor (GCSFR), a class I cytokine receptor that controls neutrophil lineage commitment, granulocytic differentiation, and survival in hematopoietic cells. Ligand-induced receptor activation engages JAK/STAT signaling—especially STAT3 and STAT5—and interfaces with MAPK/ERK and PI3K/AKT pathways to coordinate proliferation and functional maturation. Dysregulated CSF3R signaling and receptor variants have been linked to altered myeloid cell homeostasis and aberrant granulopoiesis, making this axis relevant for studies of myeloid differentiation programs and inflammatory cell dynamics. As a surface receptor with defined downstream transcriptional outputs, GCSFR is a useful node for mechanistic pathway mapping and transcriptional network analyses in immune and hematology research models.

G-CSFR CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous CSF3R expression without altering the underlying DNA sequence.

G-CSFR CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the CSF3R 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 CSF3R transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous G-CSFR expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native CSF3R locus and enabling the study of G-CSFR-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of G-CSFR pathway restoration in tumor cells with silenced or reduced CSF3R expression.

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