BNPI CRISPR Activation Plasmid (h): sc-403333-ACT

SLC17A7 encodes the vesicular glutamate transporter BNPI (VGLUT1), a synaptic vesicle membrane protein that loads glutamate into vesicles to support quantal neurotransmitter release at excitatory synapses. By controlling presynaptic glutamate packaging, BNPI influences synaptic transmission, activity-dependent plasticity, and excitation–inhibition balance across neural circuits. SLC17A7 function is closely linked to neuronal signaling pathways that shape learning and memory, including processes governing vesicle cycling, neurotransmitter homeostasis, and circuit excitability. Altered glutamatergic signaling involving SLC17A7 has been investigated in the context of neurodevelopmental and neuropsychiatric phenotypes, as well as seizure susceptibility and excitotoxic stress mechanisms.

BNPI CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous SLC17A7 expression without altering the underlying DNA sequence.

BNPI CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the SLC17A7 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 SLC17A7 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous BNPI expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native SLC17A7 locus and enabling the study of BNPI-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of BNPI pathway restoration in tumor cells with silenced or reduced SLC17A7 expression.

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