HuR/ELAV1 CRISPR Activation Plasmid (h): sc-400141-ACT

ELAVL1 (HuR) is a ubiquitously expressed RNA-binding protein that recognizes AU-rich elements in 3′ UTRs to stabilize target mRNAs and modulate translation. By coordinating post-transcriptional gene regulation, HuR influences cell-cycle progression, stress granule dynamics, DNA damage responses, and inflammatory signaling, including NF-κB- and MAPK-linked programs. HuR shuttling between nucleus and cytoplasm integrates cues from oxidative stress and cytokine signaling to reshape transcriptome output. Dysregulated ELAVL1 activity has been associated with altered control of apoptosis, senescence, and pro-oncogenic gene expression networks in multiple disease-relevant cellular contexts.

HuR/ELAV1 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous ELAVL1 expression without altering the underlying DNA sequence.

HuR/ELAV1 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the ELAVL1 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 ELAVL1 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous HuR/ELAV1 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native ELAVL1 locus and enabling the study of HuR/ELAV1-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of HuR/ELAV1 pathway restoration in tumor cells with silenced or reduced ELAVL1 expression.

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