OLIG2 CRISPR Activation Plasmid (m): sc-424530-ACT

Olig2 encodes OLIG2, a basic helix-loop-helix transcription factor that orchestrates neural lineage specification in the developing and adult mouse central nervous system. OLIG2 integrates with proneural and glial regulatory networks to control progenitor proliferation, migration, and fate decisions, with prominent roles in motor neuron and oligodendrocyte differentiation and myelin-associated gene programs. Its activity intersects with developmental signaling pathways that shape neural patterning and chromatin state, influencing transcriptional programs that define regional identity. Dysregulated OLIG2 expression and lineage programs are commonly studied in neurodevelopmental disorders, demyelinating pathology, and glioma biology as models of aberrant progenitor behavior and glial fate control.

OLIG2 CRISPR Activation Plasmid (m) provides a targeted, non-destructive approach to upregulating endogenous Olig2 expression without altering the underlying DNA sequence.

OLIG2 CRISPR Activation Plasmid (m) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the Olig2 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 Olig2 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous OLIG2 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native Olig2 locus and enabling the study of OLIG2-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of OLIG2 pathway restoration in tumor cells with silenced or reduced Olig2 expression.

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