VAChT CRISPR/Cas9 KO Plasmid (h): sc-401240
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- VAChT CRISPR/Cas9 Knockout (KO) Plasmid (h) is a pool of plasmids, each encoding Cas9 nuclease and a target-specific 20 nt guide RNA (gRNA) designed for maximum knockout efficiency using sequences derived from the GeCKO v2 library
- gRNA sequences direct Cas9 to induce site-specific double-strand breaks (DSBs) in the VAChT genomic locus, resulting in gene knockout through non-homologous end joining (NHEJ)
- The puromycin resistance and RFP genes are flanked by LoxP sites, enabling removal of selection markers via Cre recombinase (Cre Vector: sc-418923) after establishing stable knockout cell lines
- Following transfection, gene knockout efficiency can be assayed by WB, IF or IHC using antibody: VAChT Antibody (H-10): sc-518260
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
VAChT CRISPR/Cas9 KO Plasmid (h) | sc-401240 | 20 µg | $397.00 |
Overview
SLC18A3 encodes vesicular acetylcholine transporter (VAChT), a synaptic vesicle membrane protein that loads acetylcholine into secretory vesicles by exploiting the vesicular proton electrochemical gradient. This vesicular packaging step is essential for quantal cholinergic neurotransmission at neuromuscular junctions and within central and peripheral cholinergic circuits. VAChT function interfaces with acetylcholine synthesis and recycling pathways, coupling presynaptic vesicle cycling to regulated exocytosis and synaptic signaling. Altered cholinergic tone linked to disrupted acetylcholine storage has been associated with neurodevelopmental and neurodegenerative phenotypes, autonomic dysfunction, and disorders affecting motor control and cognition.
VAChT CRISPR/Cas9 KO Plasmid (h) is a pool of plasmids designed for targeted disruption of the SLC18A3 gene in human cell lines. Each plasmid co-expresses a unique single guide RNA (sgRNA) targeting a distinct site within the SLC18A3 together with the Streptococcus pyogenes Cas9 nuclease. The plasmids also encode GFP, allowing fluorescent identification and enrichment of successfully transfected cells by fluorescence microscopy or flow cytometry.
The multi-guide design increases the likelihood of generating insertions or deletions (indels) that disrupt the SLC18A3 open reading frame following Cas9-mediated double-strand break formation. DNA breaks introduced by the CRISPR/Cas9 system are repaired through endogenous non-homologous end joining (NHEJ) pathways, frequently resulting in frameshift mutations that abolish VAChT protein expression.
This CRISPR knockout system enables efficient generation of SLC18A3-deficient cell models for investigation of VAChT signaling, functional genomics studies, cancer biology research, and evaluation of therapeutic responses in human cell lines.
Key Features
- sgRNAs targeting SLC18A3 exon(s) critical for VAChT function
- Co-expression of SpCas9 and sgRNA from a single plasmid for simplified delivery
- GFP reporter for identification of transfected cells
- Pool of plasmids targeting multiple SLC18A3 genomic sites to improve knockout efficiency
- Compatible with delivery by transfection
Design Variants
CRISPRs +/- HDRs
- gRNAs encoded by VAChT CRISPR/Cas9 KO Plasmid (h) and VAChT CRISPR/Cas9 KO Plasmid (h2) target distinct sites within the SLC18A3 locus. One or both targeting designs may be available. See Related Products for availability.
- HDR donor constructs encoded by VAChT HDR Plasmid (h) and VAChT HDR Plasmid (h2) contain a puromycin resistance cassette and an RFP reporter flanked by SLC18A3 homology arms to support homology-directed repair at defined SLC18A3 target sites corresponding to the CRISPR/Cas9 KO designs. HDR donor availability may vary. See Related Products for availability.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.