Meis2 CRISPR Activation Plasmid (m): sc-421710-ACT

MEIS2 (Meis homeobox 2) is a TALE-class homeodomain transcription factor that cooperates with PBX and HOX proteins to regulate enhancer activity and lineage-specific gene expression programs during embryonic development. In mouse, Meis2 contributes to patterning and differentiation in neural crest derivatives, craniofacial morphogenesis, limb and cardiac development, and maturation of select neuronal populations, integrating cues from developmental signaling networks such as retinoic acid and HOX-regulated transcriptional circuits. By controlling cell fate specification and morphogenetic processes, altered MEIS2 dosage or dysregulated transcriptional output is relevant to congenital developmental phenotypes and neurodevelopmental biology. Meis2 is therefore frequently studied in models of differentiation, organogenesis, and transcriptional network architecture.

Meis2 CRISPR Activation Plasmid (m) provides a targeted, non-destructive approach to upregulating endogenous Meis2 expression without altering the underlying DNA sequence.

Meis2 CRISPR Activation Plasmid (m) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the Meis2 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 Meis2 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous Meis2 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native Meis2 locus and enabling the study of Meis2-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of Meis2 pathway restoration in tumor cells with silenced or reduced Meis2 expression.

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