ELL CRISPR Activation Plasmid (h): sc-404799-ACT

Human ELL encodes an RNA polymerase II elongation factor that enhances transcriptional processivity and helps coordinate productive elongation during gene expression. ELL participates in transcriptional control programs that influence cell cycle progression, differentiation, and cellular responses to stress through regulation of promoter-proximal pausing and elongation-associated checkpoints. Dysregulation of ELL-dependent transcriptional elongation is linked to altered expression of growth and lineage programs, and ELL has been implicated in oncogenic transcriptional networks through recurrent fusion events and aberrant cofactor recruitment. As a result, ELL is frequently studied for its role in transcriptional fidelity, chromatin-associated regulation, and disease-relevant gene expression remodeling.

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

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

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