TBR-1 CRISPR Activation Plasmid (h): sc-402507-ACT

TBR1 encodes the T-box transcription factor TBR-1, a nuclear regulator with essential roles in cortical neuron differentiation, neuronal migration, and the establishment of corticothalamic connectivity during human neurodevelopment. TBR-1 modulates gene expression programs downstream of developmental signaling and transcriptional networks, including interactions with factors involved in glutamatergic lineage specification and synaptic maturation. Dysregulated TBR1 expression or function has been linked to neurodevelopmental phenotypes, with recurrent associations to autism spectrum disorder and related cognitive and speech impairments through altered neuronal circuit formation. As a pathway node controlling neuronal identity and connectivity, TBR-1 is widely studied for mechanisms of transcriptional control in developing and mature neural systems.

TBR-1 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous TBR1 expression without altering the underlying DNA sequence.

TBR-1 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the TBR1 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 TBR1 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous TBR-1 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native TBR1 locus and enabling the study of TBR-1-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of TBR-1 pathway restoration in tumor cells with silenced or reduced TBR1 expression.

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