FER1L6 CRISPR Activation Plasmid (h): sc-405070-ACT

FER1L6 encodes a member of the ferlin family of large, multi-C2 domain proteins implicated in Ca2+-dependent membrane dynamics and phospholipid-dependent protein interactions. Although FER1L6 is less well characterized than other ferlins, this protein class is broadly linked to vesicle trafficking, membrane repair, endocytosis, and regulation of membrane fusion events that influence cell signaling and cytoskeletal remodeling. FER1L6 expression and genomic alterations have been reported across multiple transcriptomic and cancer-genomics datasets, suggesting potential relevance to processes such as proliferation, migration, and stress adaptation in specific cellular contexts. These features make FER1L6 a useful target for mechanistic studies of membrane-associated pathways and phenotype modulation in human cell models.

FER1L6 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous FER1L6 expression without altering the underlying DNA sequence.

FER1L6 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the FER1L6 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 FER1L6 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous FER1L6 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native FER1L6 locus and enabling the study of FER1L6-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of FER1L6 pathway restoration in tumor cells with silenced or reduced FER1L6 expression.

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