mAChR M1 CRISPR Activation Plasmid (h): sc-401453-ACT

CHRM1 encodes the human muscarinic acetylcholine receptor M1 (mAChR M1), a G protein–coupled receptor that primarily signals through Gq/11 to activate phospholipase C, elevate intracellular Ca2+, and stimulate PKC-dependent cascades. Downstream effects include modulation of MAPK/ERK signaling, phosphoinositide turnover, and activity-dependent transcription programs that shape neuronal excitability and synaptic plasticity. mAChR M1 also influences cholinergic regulation of cortical and hippocampal circuits as well as secretory and smooth muscle responses in peripheral tissues. Altered CHRM1 expression or signaling has been implicated in neuropsychiatric and neurodegenerative research contexts, including studies of cognition, attention, and cholinergic dysfunction.

mAChR M1 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous CHRM1 expression without altering the underlying DNA sequence.

mAChR M1 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the CHRM1 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 CHRM1 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous mAChR M1 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native CHRM1 locus and enabling the study of mAChR M1-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of mAChR M1 pathway restoration in tumor cells with silenced or reduced CHRM1 expression.

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