MYT1L CRISPR Activation Plasmid (h): sc-407946-ACT

MYT1L encodes a neuronal zinc-finger transcription factor that supports lineage specification and maturation programs in the developing and postnatal nervous system. MYT1L modulates chromatin and transcriptional networks that help maintain neuronal identity while repressing non-neuronal gene expression, influencing pathways linked to neurogenesis, synaptic function, and activity-dependent gene regulation. Genetic variation and dysregulated MYT1L expression have been associated with neurodevelopmental phenotypes, including autism spectrum disorder, intellectual disability, and related behavioral traits. As a regulator of neuronal transcriptional state, MYT1L is widely studied in models of neuronal differentiation, reprogramming, and circuit development.

MYT1L CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous MYT1L expression without altering the underlying DNA sequence.

MYT1L CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the MYT1L 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 MYT1L transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous MYT1L expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native MYT1L locus and enabling the study of MYT1L-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of MYT1L pathway restoration in tumor cells with silenced or reduced MYT1L expression.

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