Neurofibromin 2/NF2/Merlin CRISPR Activation Plasmid (h): sc-400504-ACT

NF2 encodes merlin (neurofibromin 2), a membrane-associated tumor suppressor that links the cortical actin cytoskeleton to transmembrane receptors and junctional complexes to regulate contact inhibition and cell mechanics. Merlin modulates Hippo pathway signaling by constraining YAP/TAZ activity and intersects with receptor tyrosine kinase, PI3K–AKT, and MAPK networks to coordinate proliferation, migration, and polarity. It also contributes to adherens and tight junction stability, influencing tissue architecture and mechanotransduction responses. Disruption of NF2 function is implicated in neurofibromatosis type 2 and a spectrum of merlin-deficient tumors, making NF2 a key node for studying growth control and cytoskeletal signaling.

Neurofibromin 2/NF2/Merlin CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous NF2 expression without altering the underlying DNA sequence.

Neurofibromin 2/NF2/Merlin CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the NF2 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 NF2 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous Neurofibromin 2/NF2/Merlin expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native NF2 locus and enabling the study of Neurofibromin 2/NF2/Merlin-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of Neurofibromin 2/NF2/Merlin pathway restoration in tumor cells with silenced or reduced NF2 expression.

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