Klotho CRISPR Activation Plasmid (m): sc-421290-ACT

Mouse Kl encodes Klotho, a type I membrane protein that can be shed to form a circulating factor with broad effects on mineral metabolism and tissue homeostasis. Klotho functions as an obligate co-receptor for endocrine FGF signaling, particularly FGF23, coordinating phosphate handling, vitamin D metabolism, and calcium balance. Beyond its endocrine role, Klotho has been linked to modulation of oxidative stress responses and cellular senescence-associated programs that influence kidney, cardiovascular, and nervous system physiology. Altered Klotho expression is frequently studied in models of chronic kidney injury, age-associated functional decline, and metabolic dysregulation to clarify pathway-level mechanisms.

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

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

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