LHX9 CRISPR Activation Plasmid (h): sc-403360-ACT

LHX9 (LIM homeobox 9) encodes a LIM-domain transcription factor that regulates cell fate decisions during embryonic development by controlling lineage-specific gene expression programs. In human biology, LHX9 activity is linked to patterning and differentiation processes in the developing nervous system and gonadal tissues, where precise temporal transcriptional control is required. As a DNA-binding regulator, LHX9 interfaces with chromatin and transcriptional networks that shape developmental pathways, and altered expression has been investigated in contexts of dysregulated differentiation and oncogenic transcriptional states. These properties make LHX9 a useful target for studying gene regulatory circuitry, developmental transcription factors, and phenotype modulation driven by changes in endogenous transcription.

LHX9 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous LHX9 expression without altering the underlying DNA sequence.

LHX9 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the LHX9 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 LHX9 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous LHX9 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native LHX9 locus and enabling the study of LHX9-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of LHX9 pathway restoration in tumor cells with silenced or reduced LHX9 expression.

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