ferritin light chain CRISPR Activation Plasmid (h): sc-400924-ACT

FTL encodes ferritin light chain, a core component of the ferritin nanocage that buffers intracellular labile iron by storing Fe(III) and limiting iron-catalyzed reactive oxygen species generation. By coordinating iron sequestration with ferritin heavy chain activity and ferritinophagy, FTL helps regulate redox balance, mitochondrial function, and cellular responses to oxidative stress. Altered ferritin composition or iron handling influences pathways linked to ferroptosis susceptibility and can modulate inflammatory and metabolic stress signaling. Dysregulated FTL expression has been associated with iron overload states and neurodegenerative and hematologic phenotypes where iron homeostasis and oxidative damage are prominent biological features.

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

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

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