BETA 3 CRISPR Activation Plasmid (m2): sc-425594-ACT-2

Mouse Bhlhe22 encodes the basic helix–loop–helix transcription factor BETA3, a regulator of neuronal differentiation and maturation that helps shape gene expression programs during central nervous system development. BETA3 modulates transcriptional networks linked to cell fate specification, neurite outgrowth, and synaptic organization through bHLH-mediated DNA binding and interaction with developmental signaling pathways that coordinate progenitor-to-neuron transitions. Altered Bhlhe22 activity has been associated in the literature with disrupted cortical circuitry and neurodevelopmental phenotypes, making it relevant for studying mechanisms underlying brain development and neurological disease biology. Gene editing of Bhlhe22 in mouse models supports functional interrogation of lineage decisions, circuit assembly, and downstream transcriptomic changes in primary neurons, organoids, or in vivo developmental contexts.

BETA 3 CRISPR Activation Plasmid (m2) provides a targeted, non-destructive approach to upregulating endogenous Bhlhe22 expression without altering the underlying DNA sequence.

BETA 3 CRISPR Activation Plasmid (m2) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the Bhlhe22 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 Bhlhe22 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous BETA 3 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native Bhlhe22 locus and enabling the study of BETA 3-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of BETA 3 pathway restoration in tumor cells with silenced or reduced Bhlhe22 expression.

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