c-Mpl CRISPR Activation Plasmid (h): sc-403035-ACT

MPL encodes the thrombopoietin receptor c-Mpl, a class I cytokine receptor that regulates hematopoietic stem and progenitor cell maintenance and drives megakaryocyte proliferation and platelet production. Upon thrombopoietin binding, c-Mpl signals through JAK2/STAT5, MAPK/ERK, and PI3K/AKT pathways to coordinate survival, differentiation, and cytokine-responsive transcriptional programs. Dysregulated MPL signaling or expression is linked to aberrant hematopoiesis and is frequently studied in the context of myeloproliferative neoplasms and inherited thrombocytopenia syndromes. As a surface receptor with well-defined downstream readouts, c-Mpl provides a tractable node for interrogating cytokine signaling, lineage commitment, and stress hematopoiesis in human cell models.

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

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

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