Sp4 CRISPR Activation Plasmid (h): sc-402568-ACT

Human SP4 encodes the Sp4 transcription factor, a member of the Sp/KLF family that binds GC-rich promoter elements to regulate gene expression programs controlling neuronal differentiation, synaptic plasticity, and activity-dependent transcription. Sp4 contributes to chromatin- and transcriptional control processes that shape neurodevelopmental trajectories and neuronal excitability, linking it to pathways governing axon/dendrite maturation and circuit formation. Altered SP4/Sp4 expression or function has been associated with neuropsychiatric and neurodevelopmental phenotypes, supporting its relevance for studying transcriptional network dysregulation in the nervous system. In cell models, modulating SP4 can be used to interrogate downstream target genes and gene regulatory circuitry influencing neuronal and glial states.

Sp4 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous SP4 expression without altering the underlying DNA sequence.

Sp4 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the SP4 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 SP4 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous Sp4 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native SP4 locus and enabling the study of Sp4-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of Sp4 pathway restoration in tumor cells with silenced or reduced SP4 expression.

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