INTS3 CRISPR Activation Plasmid (h): sc-408952-ACT

Integrator complex subunit 3 (INTS3) is a core component of the Integrator complex, a multiprotein assembly that couples RNA polymerase II transcription with 3′ end processing of small nuclear RNAs and broader regulation of transcriptional output. INTS3 also participates in genome maintenance programs through interactions linked to DNA damage sensing and repair, supporting replication stress responses and preservation of chromosomal integrity. By shaping RNA processing and transcriptional homeostasis, INTS3 influences cell-cycle progression and stress-adaptive gene expression networks. Dysregulation of Integrator-associated pathways and DNA repair coordination is implicated in altered proliferation and genome instability phenotypes relevant to cancer biology and other disorders of nuclear RNA metabolism.

INTS3 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous INTS3 expression without altering the underlying DNA sequence.

INTS3 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the INTS3 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 INTS3 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous INTS3 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native INTS3 locus and enabling the study of INTS3-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of INTS3 pathway restoration in tumor cells with silenced or reduced INTS3 expression.

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