LHX2 CRISPR Activation Plasmid (h): sc-401071-ACT

LHX2 (LIM homeobox 2) is a nuclear transcription factor containing LIM domains and a homeodomain that regulates cell fate decisions during development and tissue patterning. In human cells, LHX2 contributes to transcriptional programs controlling proliferation, differentiation, and maintenance of progenitor-like states, with downstream effects on chromatin organization and lineage-specifying gene networks. Its activity intersects with developmental signaling and transcriptional regulatory circuitry that shapes organogenesis, including neural and epithelial contexts. Dysregulated LHX2 expression has been reported in multiple disease-associated expression signatures, making it a useful node for studying mechanisms of aberrant differentiation and transcriptional control.

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

LHX2 CRISPR Activation Plasmid (h) 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.