betaKlotho CRISPR Activation Plasmid (h): sc-401614-ACT

Human KLB encodes betaKlotho, a single-pass transmembrane co-receptor that confers ligand specificity to endocrine FGF signaling, most notably facilitating FGF19/FGFR4 signaling in hepatocytes and FGF21/FGFR1c signaling in metabolic tissues. Through these pathways, betaKlotho regulates bile acid homeostasis, glucose and lipid metabolism, and energy balance by modulating downstream MAPK/ERK and related transcriptional programs. Altered KLB expression or function has been linked to metabolic dysregulation, including obesity, insulin resistance, and nonalcoholic fatty liver disease–associated processes, and may influence hepatic growth and stress responses. Because KLB is a key determinant of endocrine FGF responsiveness, it is frequently studied in models of nutrient sensing, hepatocyte signaling, and adipose–liver crosstalk.

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

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

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