SCML2 CRISPR Activation Plasmid (h): sc-405003-ACT

SCML2 (sex comb on midleg-like 2) encodes a Polycomb group protein that contributes to epigenetic gene silencing through chromatin-associated complexes and histone mark–dependent regulation. It is linked to transcriptional repression programs that shape cell identity, differentiation, and germline and neural developmental processes, consistent with its chromatin reader functions and interactions with Polycomb regulatory machinery. By modulating accessibility at developmental loci, SCML2 influences genome-wide transcriptional states and can intersect with pathways controlling cell-cycle progression and lineage commitment. Dysregulated SCML2 expression or altered Polycomb activity has been associated with aberrant epigenetic states observed in cancer and neurodevelopmental phenotypes, making it relevant for studies of transcriptional control and disease-associated chromatin remodeling.

SCML2 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous SCML2 expression without altering the underlying DNA sequence.

SCML2 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the SCML2 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 SCML2 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous SCML2 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native SCML2 locus and enabling the study of SCML2-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of SCML2 pathway restoration in tumor cells with silenced or reduced SCML2 expression.

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