M-CSF CRISPR Activation Plasmid (h): sc-401339-ACT

CSF1 encodes macrophage colony-stimulating factor (M-CSF), a secreted cytokine that governs survival, proliferation, and differentiation of monocytes and macrophages through CSF1R signaling. Ligand-dependent receptor activation engages PI3K–AKT, MAPK/ERK, and JAK/STAT pathways to regulate myeloid lineage commitment, phagocytic function, and inflammatory gene expression. In human tissues, M-CSF contributes to osteoclastogenesis and bone remodeling, shaping immune–stromal crosstalk in the hematopoietic and skeletal microenvironments. Dysregulated CSF1/M-CSF signaling has been implicated in altered macrophage polarization and myeloid infiltration patterns observed across inflammatory and neoplastic contexts, supporting its use as a mechanistic node in immunology and tumor microenvironment research.

M-CSF CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous CSF1 expression without altering the underlying DNA sequence.

M-CSF CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the CSF1 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 CSF1 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous M-CSF expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native CSF1 locus and enabling the study of M-CSF-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of M-CSF pathway restoration in tumor cells with silenced or reduced CSF1 expression.

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