RUNX1 CRISPR Activation Plasmid (m): sc-419483-ACT

Runx1 encodes the Runt-related transcription factor RUNX1, a core regulator of definitive hematopoiesis that controls lineage commitment and maturation of hematopoietic stem and progenitor cells. RUNX1 operates in transcriptional networks with cofactors such as CBFβ and integrates signaling inputs to coordinate chromatin accessibility, enhancer activity, and cell-cycle programs during myeloid and lymphoid differentiation. In mouse models, altered Runx1 dosage or activity perturbs blood development, impacts immune cell homeostasis, and provides mechanistic links to leukemogenesis and bone marrow failure phenotypes. This gene is frequently studied in pathways governing stem cell self-renewal, differentiation checkpoints, and oncogenic transcriptional dysregulation.

RUNX1 CRISPR Activation Plasmid (m) provides a targeted, non-destructive approach to upregulating endogenous Runx1 expression without altering the underlying DNA sequence.

RUNX1 CRISPR Activation Plasmid (m) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the Runx1 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 Runx1 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous RUNX1 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native Runx1 locus and enabling the study of RUNX1-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of RUNX1 pathway restoration in tumor cells with silenced or reduced Runx1 expression.

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