FRRS1L CRISPR Activation Plasmid (h): sc-407611-ACT

FRRS1L (ferric chelate reductase 1 like) encodes an endoplasmic reticulum–associated neuronal protein implicated in the maturation and functional regulation of AMPA-type glutamate receptors. By supporting receptor assembly/trafficking and synaptic localization, FRRS1L contributes to excitatory neurotransmission, synaptic plasticity, and activity-dependent signaling networks in the central nervous system. Disruption of FRRS1L function has been linked to neurodevelopmental and epileptic phenotypes, consistent with impaired glutamatergic signaling and altered neuronal network excitability. These features make FRRS1L a useful target for studying synapse biology, receptor proteostasis, and pathways governing neuronal development and circuit function.

FRRS1L CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous FRRS1L expression without altering the underlying DNA sequence.

FRRS1L CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the FRRS1L 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 FRRS1L transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous FRRS1L expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native FRRS1L locus and enabling the study of FRRS1L-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of FRRS1L pathway restoration in tumor cells with silenced or reduced FRRS1L expression.

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