HLX1 CRISPR Activation Plasmid (h): sc-403700-ACT

Human HLX (HLX1) encodes a homeobox transcription factor that regulates cell fate decisions during embryonic development and hematopoietic lineage specification. HLX1 modulates transcriptional programs controlling proliferation, differentiation, and tissue patterning, with downstream effects on developmental signaling networks and immune cell maturation. Dysregulated HLX expression has been reported in hematologic and solid tumor contexts, where altered transcriptional control can impact oncogenic signaling, differentiation state, and cellular invasiveness. These properties make HLX1 a useful node for dissecting transcriptional circuitry linking development, immune biology, and disease-associated gene expression programs.

HLX1 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous HLX expression without altering the underlying DNA sequence.

HLX1 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the HLX 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 HLX transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous HLX1 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native HLX locus and enabling the study of HLX1-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of HLX1 pathway restoration in tumor cells with silenced or reduced HLX expression.

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