NuMA CRISPR Activation Plasmid (h): sc-401570-ACT

Human NUMA1 encodes nuclear mitotic apparatus protein (NuMA), a large coiled-coil scaffold that concentrates at spindle poles to organize microtubule minus ends and ensure accurate chromosome segregation during mitosis. NuMA cooperates with the dynein–dynactin complex and LGN/Gαi-mediated cortical pathways to position the spindle, support spindle pole integrity, and coordinate cytokinesis and nuclear architecture. Perturbation of NUMA1 function can disrupt mitotic fidelity, promote aneuploidy, and alter cell polarity decisions, making it a relevant node for studies of genome stability and proliferation-associated phenotypes. These properties link NuMA to mechanistic investigations in cancer biology and developmental cell biology where spindle orientation and chromosomal stability are central.

NuMA CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous NUMA1 expression without altering the underlying DNA sequence.

NuMA CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the NUMA1 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 NUMA1 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous NuMA expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native NUMA1 locus and enabling the study of NuMA-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of NuMA pathway restoration in tumor cells with silenced or reduced NUMA1 expression.

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