Id1 CRISPR Activation Plasmid (h): sc-400341-ACT

Human ID1 encodes Id1, a helix–loop–helix (HLH) transcriptional regulator that lacks a DNA-binding domain and modulates gene expression by sequestering E-proteins and other bHLH factors. By restraining lineage-specific transcriptional programs, Id1 helps control proliferation, differentiation, and cellular plasticity, and it is frequently regulated downstream of BMP/TGF-β family signaling and related SMAD-dependent transcriptional networks. ID1 activity is linked to developmental and stem-like states, with context-dependent roles in cell-cycle progression, migration, and stress responses. Dysregulated ID1 expression is reported across multiple disease-relevant models, where altered differentiation and signaling balance can contribute to aberrant tissue remodeling and oncogenic phenotypes.

Id1 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous ID1 expression without altering the underlying DNA sequence.

Id1 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the ID1 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 ID1 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous Id1 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native ID1 locus and enabling the study of Id1-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of Id1 pathway restoration in tumor cells with silenced or reduced ID1 expression.

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