LTBP-3 CRISPR Activation Plasmid (h): sc-404036-ACT

LTBP3 encodes latent transforming growth factor beta binding protein 3 (LTBP-3), an extracellular matrix glycoprotein that binds the latent TGF-β complex and helps control its sequestration, storage, and spatial activation in tissues. By regulating TGF-β bioavailability, LTBP-3 contributes to SMAD-dependent signaling and extracellular matrix remodeling processes that influence cell adhesion, migration, differentiation, and tissue homeostasis. Altered LTBP3 expression or function has been associated with dysregulated TGF-β signaling in contexts including fibrosis, connective tissue remodeling, and tumor microenvironment biology, making it relevant for mechanistic studies of matrix-dependent signaling. Researchers commonly interrogate LTBP-3 in pathways linking ECM organization to growth factor activation and downstream transcriptional programs.

LTBP-3 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous LTBP3 expression without altering the underlying DNA sequence.

LTBP-3 CRISPR Activation Plasmid (h) 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.