LTBP-3 CRISPR Activation Plasmid (m): sc-421482-ACT

Ltbp3 encodes latent transforming growth factor beta binding protein 3 (LTBP-3), an extracellular matrix glycoprotein that binds latent TGF-β complexes and regulates their sequestration, activation, and spatial presentation in the tissue microenvironment. By controlling TGF-β bioavailability, LTBP-3 influences SMAD-dependent signaling, matrix remodeling, and cell fate decisions during development and tissue homeostasis. Mouse studies link LTBP-3 to skeletal patterning, connective tissue integrity, and vascular biology, reflecting its role in coordinating growth factor signaling with extracellular matrix organization. Dysregulated LTBP-3–TGF-β axis activity is relevant to fibrosis-like remodeling, aberrant osteogenesis, and tumor-associated stromal signaling in mechanistic research settings.

LTBP-3 CRISPR Activation Plasmid (m) provides a targeted, non-destructive approach to upregulating endogenous Ltbp3 expression without altering the underlying DNA sequence.

LTBP-3 CRISPR Activation Plasmid (m) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the Ltbp3 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 Ltbp3 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous LTBP-3 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native Ltbp3 locus and enabling the study of LTBP-3-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of LTBP-3 pathway restoration in tumor cells with silenced or reduced Ltbp3 expression.

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