Chitotriosidase CRISPR Activation Plasmid (h): sc-402112-ACT

CHIT1 encodes chitotriosidase, a secreted and lysosomal glycoside hydrolase predominantly expressed by activated macrophages and other myeloid cells. Although humans do not synthesize chitin, CHIT1 participates in innate immune responses by recognizing and degrading chitin-containing structures from fungi and parasites and is linked to macrophage polarization, lysosomal function, and inflammatory signaling programs. Altered CHIT1 expression and enzymatic activity have been associated with disorders featuring macrophage activation and tissue remodeling, including lysosomal storage diseases and chronic inflammatory conditions. As a readout of myeloid activation, CHIT1 is frequently studied in contexts of host–pathogen interactions, granulomatous inflammation, and fibrotic microenvironments.

Chitotriosidase CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous CHIT1 expression without altering the underlying DNA sequence.

Chitotriosidase CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the CHIT1 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 CHIT1 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous Chitotriosidase expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native CHIT1 locus and enabling the study of Chitotriosidase-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of Chitotriosidase pathway restoration in tumor cells with silenced or reduced CHIT1 expression.

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