FMR1 CRISPR Activation Plasmid (h): sc-401919-ACT

FMR1 encodes fragile X mental retardation protein (FMRP), an RNA-binding protein that regulates mRNA transport, stability, and translational repression at synapses and in other neuronal compartments. FMRP modulates activity-dependent protein synthesis through interactions with ribonucleoprotein complexes and signaling pathways that shape synaptic plasticity, including mGluR-dependent translation and broader neuronal development programs. Dysregulation of FMR1 expression or function is linked to fragile X–associated disorders and neurodevelopmental phenotypes, making it a widely used node for studying post-transcriptional control in the brain. In cultured cells and model systems, FMR1 is commonly investigated to map RNA targets, define translational control networks, and interrogate mechanisms of activity-responsive gene expression.

FMR1 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous FMR1 expression without altering the underlying DNA sequence.

FMR1 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the FMR1 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 FMR1 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous FMR1 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native FMR1 locus and enabling the study of FMR1-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of FMR1 pathway restoration in tumor cells with silenced or reduced FMR1 expression.

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