G-CSF CRISPR Activation Plasmid (m): sc-419844-ACT

Mouse Csf3 encodes granulocyte colony-stimulating factor (G-CSF), a secreted cytokine that controls granulopoiesis by promoting proliferation, differentiation, and survival of neutrophil progenitors and by mobilizing mature granulocytes from bone marrow. G-CSF signals primarily through CSF3R to activate JAK/STAT, MAPK/ERK, and PI3K/AKT pathways, coordinating innate immune homeostasis and inflammatory responses. Altered Csf3/G-CSF axis activity is commonly studied in models of infection and sterile inflammation, as well as in settings of myeloid expansion and neutrophil-driven tissue injury. In tumor biology and immunology, G-CSF is also used to probe how cytokine-driven myelopoiesis shapes the myeloid compartment within inflammatory microenvironments.

G-CSF CRISPR Activation Plasmid (m) provides a targeted, non-destructive approach to upregulating endogenous Csf3 expression without altering the underlying DNA sequence.

G-CSF CRISPR Activation Plasmid (m) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the Csf3 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 Csf3 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous G-CSF expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native Csf3 locus and enabling the study of G-CSF-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of G-CSF pathway restoration in tumor cells with silenced or reduced Csf3 expression.

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