METRNL CRISPR Activation Plasmid (m): sc-431669-ACT

Metrnl encodes meteorin-like (METRNL), a secreted protein implicated in intercellular communication that coordinates metabolic homeostasis and immunomodulatory programs in peripheral tissues. In mouse models, METRNL expression is linked to adipose tissue remodeling, macrophage polarization, and adaptive thermogenic responses, integrating nutrient sensing with inflammatory signaling. Reported activities connect METRNL to pathways involved in cytokine signaling, extracellular matrix interactions, and tissue repair processes that influence systemic energy balance. Dysregulation of Metrnl has been investigated in the context of obesity-associated inflammation, insulin resistance, and cardiometabolic phenotypes, supporting its utility as a node for mechanistic studies of metabolic disease biology.

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

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

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