Dyrk1A CRISPR Activation Plasmid (h): sc-401928-ACT

DYRK1A encodes dual-specificity tyrosine phosphorylation-regulated kinase 1A (Dyrk1A), a serine/threonine kinase that modulates protein phosphorylation programs governing cell cycle progression, neuronal differentiation, and synaptic function. Dyrk1A integrates with signaling networks that control transcriptional regulation, RNA splicing, and proteostasis, with reported substrates and pathway links that influence chromatin state and stress responses. Altered DYRK1A dosage and activity are implicated in neurodevelopmental phenotypes, including dosage-sensitive effects associated with Down syndrome and autism spectrum–related biology, and have also been studied in contexts of cancer cell proliferation. These features make DYRK1A a useful node for dissecting kinase-driven regulatory circuits in human cellular models.

Dyrk1A CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous DYRK1A expression without altering the underlying DNA sequence.

Dyrk1A CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the DYRK1A 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 DYRK1A transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous Dyrk1A expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native DYRK1A locus and enabling the study of Dyrk1A-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of Dyrk1A pathway restoration in tumor cells with silenced or reduced DYRK1A expression.

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