MESP1 CRISPR Activation Plasmid (h): sc-404176-ACT

MESP1 (mesoderm posterior basic helix-loop-helix transcription factor 1) is a master regulator of early mesendoderm specification and cardiovascular lineage commitment in human development. As a bHLH transcription factor, MESP1 coordinates transcriptional programs that govern gastrulation-associated cell fate decisions, epithelial-to-mesenchymal transition, and progenitor migration, acting upstream of networks that pattern mesoderm and initiate cardiogenesis. Dysregulated MESP1 activity can perturb differentiation trajectories and has been implicated in developmental defects and altered lineage plasticity relevant to congenital heart biology and stem cell disease modeling. In vitro, controlled modulation of MESP1 supports mechanistic studies of early developmental gene regulatory networks and differentiation protocols for mesoderm-derived cell types.

MESP1 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous MESP1 expression without altering the underlying DNA sequence.

MESP1 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the MESP1 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 MESP1 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous MESP1 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native MESP1 locus and enabling the study of MESP1-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of MESP1 pathway restoration in tumor cells with silenced or reduced MESP1 expression.

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