DTX3L CRISPR Activation Plasmid (h): sc-403953-ACT

DTX3L (deltex E3 ubiquitin ligase 3L) encodes a human E3 ubiquitin ligase that participates in ubiquitin-dependent protein regulation and coordinates with PARP9 in interferon-stimulated signaling. Through modulation of ubiquitination and crosstalk with DNA damage response machinery, DTX3L contributes to control of protein turnover, chromatin-associated processes, and innate immune pathway outputs. Altered DTX3L activity has been linked to dysregulated inflammatory signaling and changes in cellular stress responses, making it relevant for studies of immune regulation and genome stability. Its molecular functions position DTX3L as a useful node for dissecting ubiquitin pathway dynamics and interferon-driven transcriptional programs.

DTX3L CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous DTX3L expression without altering the underlying DNA sequence.

DTX3L CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the DTX3L 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 DTX3L transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous DTX3L expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native DTX3L locus and enabling the study of DTX3L-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of DTX3L pathway restoration in tumor cells with silenced or reduced DTX3L expression.

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