LYRM4 CRISPR Activation Plasmid (h): sc-412759-ACT

Human LYRM4 (LYR motif containing 4), also known as ISD11, is a mitochondrial matrix protein that functions as an essential cofactor of the NFS1 cysteine desulfurase complex required for iron–sulfur (Fe–S) cluster biogenesis. By stabilizing the core sulfur transfer machinery and supporting scaffold-dependent Fe–S assembly, LYRM4 helps maintain the activity of Fe–S–dependent enzymes involved in oxidative phosphorylation, mitochondrial translation, and redox homeostasis. Disruption or insufficiency of this pathway can impair respiratory chain function, elevate oxidative stress, and compromise cellular energy metabolism, linking LYRM4-associated mitochondrial dysfunction to neurometabolic disease mechanisms. As a result, LYRM4 is frequently studied in the context of mitochondrial quality control, metabolic adaptation, and genotype-to-phenotype relationships in Fe–S cluster assembly defects.

LYRM4 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous LYRM4 expression without altering the underlying DNA sequence.

LYRM4 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the LYRM4 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 LYRM4 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous LYRM4 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native LYRM4 locus and enabling the study of LYRM4-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of LYRM4 pathway restoration in tumor cells with silenced or reduced LYRM4 expression.

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