Islet-1 CRISPR Activation Plasmid (h): sc-401733-ACT

Human ISL1 encodes Islet-1, a LIM homeobox transcription factor that functions as a lineage-specifying regulator during embryonic development. ISL1 integrates with core transcriptional programs and chromatin-regulatory networks to control gene expression in pancreatic endocrine progenitors, motor neurons, and cardiac second heart field derivatives, influencing differentiation, migration, and maturation. Through coordination with pathways governing organogenesis and cell fate decisions, ISL1 helps define endocrine identity and neurogenic programs in a context-dependent manner. Dysregulated ISL1 expression or altered downstream circuitry has been associated with developmental abnormalities and is frequently used as a molecular marker and mechanistic node in studies of diabetes-related beta-cell biology, cardiogenesis, and neurodevelopmental processes.

Islet-1 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous ISL1 expression without altering the underlying DNA sequence.

Islet-1 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the ISL1 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 ISL1 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous Islet-1 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native ISL1 locus and enabling the study of Islet-1-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of Islet-1 pathway restoration in tumor cells with silenced or reduced ISL1 expression.

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