twist CRISPR Activation Plasmid (h): sc-414117-ACT

Human TWIST2 encodes twist, a basic helix–loop–helix transcription factor that regulates lineage commitment, mesenchymal differentiation, and tissue morphogenesis. twist integrates developmental signaling with transcriptional programs controlling cell identity, extracellular matrix remodeling, and epithelial–mesenchymal transition-related processes. It modulates gene networks involved in craniofacial and limb development and influences fibroblast and immune-related transcriptional states. Dysregulated TWIST2 activity has been associated with congenital developmental syndromes and altered stromal biology, making it a useful node for studying gene regulatory circuits in development and disease-relevant cell models.

twist2 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous TWIST2 expression without altering the underlying DNA sequence.

twist2 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the TWIST2 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 TWIST2 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous twist2 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native TWIST2 locus and enabling the study of twist2-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of twist2 pathway restoration in tumor cells with silenced or reduced TWIST2 expression.

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