TLX3 CRISPR Activation Plasmid (h): sc-405569-ACT

TLX3 (T-cell leukemia homeobox 3) is a homeobox transcription factor that regulates context-dependent gene expression programs during development and lineage specification, with prominent roles in hematopoietic and neuronal differentiation. By binding specific DNA motifs through its homeodomain, TLX3 can rewire transcriptional networks that influence cell fate decisions, proliferation control, and maturation state. Dysregulated TLX3 expression is associated with altered differentiation programs and aberrant transcriptional signaling observed in hematologic malignancy models, making it a useful node for studying oncogenic transcription factor biology. In cellular systems, TLX3 perturbation is commonly interrogated to map downstream target genes, epigenetic states, and pathway crosstalk governing developmental and disease-relevant phenotypes.

TLX3 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous TLX3 expression without altering the underlying DNA sequence.

TLX3 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the TLX3 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 TLX3 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous TLX3 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native TLX3 locus and enabling the study of TLX3-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of TLX3 pathway restoration in tumor cells with silenced or reduced TLX3 expression.

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