MRTF-B CRISPR Activation Plasmid (h): sc-401793-ACT

Human MKL2 encodes myocardin-related transcription factor B (MRTF-B), a RhoA–actin dynamics–responsive coactivator that partners with SRF to regulate cytoskeletal gene programs controlling cell shape, migration, and contractility. MRTF-B activity integrates signals from actin polymerization and mechanotransduction to modulate transcriptional networks involved in smooth muscle and cardiovascular development as well as tissue remodeling. Dysregulated MKL2/MRTF-B–SRF signaling has been linked to aberrant epithelial–mesenchymal transition, invasive behavior, and fibroproliferative responses in multiple model systems. As a node connecting actin remodeling to transcriptional output, MRTF-B is widely studied in pathways governing adhesion, stress fiber formation, and extracellular matrix regulation.

MRTF-B CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous MKL2 expression without altering the underlying DNA sequence.

MRTF-B CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the MKL2 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 MKL2 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous MRTF-B expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native MKL2 locus and enabling the study of MRTF-B-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of MRTF-B pathway restoration in tumor cells with silenced or reduced MKL2 expression.

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