C12orf57 CRISPR Activation Plasmid (h): sc-408545-ACT

C12orf57 encodes a small, poorly characterized human protein that has been linked to fundamental cellular homeostasis, with emerging evidence implicating roles in neuronal development and stress-responsive processes. Although its molecular partners and pathway placement remain under active study, altered C12orf57 dosage has been associated with neurodevelopmental phenotypes and syndromic disease presentations, supporting a requirement for proper gene regulation in differentiated tissues. Transcriptional control of C12orf57 may intersect with broader programs governing cell fate decisions, proteostasis, and intracellular signaling dynamics. These features make C12orf57 a useful locus for investigating how subtle changes in expression influence cellular phenotypes relevant to neurological and developmental biology.

C12orf57 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous C12orf57 expression without altering the underlying DNA sequence.

C12orf57 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the C12orf57 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 C12orf57 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous C12orf57 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native C12orf57 locus and enabling the study of C12orf57-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of C12orf57 pathway restoration in tumor cells with silenced or reduced C12orf57 expression.

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