T2R38 CRISPR Activation Plasmid (h): sc-402279-ACT

TAS2R38 encodes the bitter taste receptor T2R38, a GPCR that detects thiourea-containing ligands and signals through canonical gustatory pathways involving G protein–coupled activation of phospholipase Cβ2, intracellular Ca²⁺ mobilization, and downstream TRPM5-dependent depolarization. Beyond oral taste perception, T2R38 is expressed in extraoral epithelia where it can influence chemosensory signaling and innate defense–associated cellular programs. Common coding polymorphisms alter receptor responsiveness and have been widely used to study inter-individual differences in bitter perception and chemosensory biology. These features make TAS2R38 a useful model for investigating GPCR signaling dynamics, epithelial sensing pathways, and genotype–phenotype relationships.

T2R38 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous TAS2R38 expression without altering the underlying DNA sequence.

T2R38 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the TAS2R38 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 TAS2R38 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous T2R38 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native TAS2R38 locus and enabling the study of T2R38-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of T2R38 pathway restoration in tumor cells with silenced or reduced TAS2R38 expression.

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