Tau CRISPR Activation Plasmid (h): sc-400136-ACT

Human MAPT encodes the microtubule-associated protein tau, a predominantly neuronal factor that binds and stabilizes microtubules to support axonal transport, neurite outgrowth, and cytoskeletal organization. Tau function is regulated by post-translational modifications, particularly phosphorylation, which modulates microtubule affinity and influences cytoskeletal dynamics and intracellular trafficking. Dysregulation of MAPT expression and tau homeostasis is linked to neurodegenerative processes and protein aggregation observed in tauopathies, including Alzheimer’s disease and frontotemporal dementia. MAPT is therefore widely studied in pathways controlling microtubule stability, synaptic integrity, proteostasis, and neuroinflammation-associated stress responses.

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

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

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