CNTF CRISPR Activation Plasmid (h): sc-402471-ACT

Human CNTF (ciliary neurotrophic factor) is a secreted cytokine in the IL-6 family that supports neuronal survival, differentiation, and stress responses in the central and peripheral nervous systems. CNTF signals primarily through the CNTFRα/LIFR/gp130 receptor complex to activate JAK/STAT pathways, with additional engagement of MAPK/ERK and PI3K/AKT signaling that influences neurite outgrowth, glial biology, and inflammatory crosstalk. Altered CNTF expression and pathway activity have been studied in neurodegeneration, retinal injury, and demyelinating or neuropathic contexts where trophic support and reactive gliosis shape disease-associated phenotypes. In vitro and in vivo models use CNTF perturbation to interrogate neuroprotection programs, neuron–glia interactions, and transcriptional networks linked to regeneration and metabolic stress.

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

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

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