LHX2 CRISPR Activation Plasmid (m): sc-421425-ACT

Mouse Lhx2 encodes the LIM homeobox transcription factor LHX2, a nuclear regulator that controls cell fate specification, tissue patterning, and maintenance of progenitor competence during embryonic and postnatal development. LHX2 participates in transcriptional programs that shape neurogenesis and cortical arealization, support retinal development, and influence epithelial–mesenchymal state decisions through context-dependent interaction with co-factors and chromatin regulators. In stem and progenitor compartments, LHX2 activity contributes to lineage restriction, differentiation timing, and survival, linking it to pathways governing developmental transcription networks and chromatin accessibility. Dysregulated LHX2 expression has been associated with developmental defects and altered proliferative states in disease-relevant models, making it a useful node for dissecting gene regulatory circuits in neuroscience and developmental biology.

LHX2 CRISPR Activation Plasmid (m) provides a targeted, non-destructive approach to upregulating endogenous Lhx2 expression without altering the underlying DNA sequence.

LHX2 CRISPR Activation Plasmid (m) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the Lhx2 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 Lhx2 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous LHX2 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native Lhx2 locus and enabling the study of LHX2-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of LHX2 pathway restoration in tumor cells with silenced or reduced Lhx2 expression.

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