Flt-3/Flk-2 CRISPR Activation Plasmid (h): sc-400218-ACT

FLT3 (Flt-3/Flk-2) encodes a class III receptor tyrosine kinase predominantly expressed in hematopoietic progenitors, where binding of FLT3 ligand promotes proliferation, survival, and differentiation. Receptor activation triggers downstream PI3K–AKT, RAS–MAPK, and JAK–STAT signaling, integrating cues that regulate myeloid and lymphoid development. Dysregulated FLT3 signaling, including activating mutations and aberrant expression, is frequently studied in the context of leukemogenesis and altered hematopoietic cell fate. As a result, FLT3 serves as a widely used molecular handle to interrogate growth factor receptor signaling, progenitor biology, and oncogenic kinase-driven network rewiring in human cell models.

Flt-3/Flk-2 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous FLT3 expression without altering the underlying DNA sequence.

Flt-3/Flk-2 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the FLT3 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 FLT3 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous Flt-3/Flk-2 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native FLT3 locus and enabling the study of Flt-3/Flk-2-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of Flt-3/Flk-2 pathway restoration in tumor cells with silenced or reduced FLT3 expression.

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