Orexin-A CRISPR/Cas9 KO Plasmid (h): sc-400357
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- Orexin-A 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 Orexin-A 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: Orexin-A Antibody (KK09): sc-80263
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
Orexin-A CRISPR/Cas9 KO Plasmid (h) | sc-400357 | 20 µg | $397.00 |
Overview
HCRT encodes prepro-orexin, which is processed to the neuropeptides orexin-A (hypocretin-1) and orexin-B that act through OX1R/HCRTR1 and OX2R/HCRTR2 GPCR signaling. Orexin-A is a key modulator of arousal and vigilance, coupling neuronal metabolic state to synaptic excitability via cAMP/PKA-, Ca2+-dependent, and MAPK/ERK-linked pathways in hypothalamic and brainstem circuits. In addition to regulating sleep–wake transitions, orexin signaling influences feeding behavior, reward processing, stress responses, and autonomic output. Dysregulation of the HCRT/orexin axis is associated with sleep–wake disorders and contributes to neurobehavioral phenotypes relevant to metabolic and psychiatric research.
Orexin-A CRISPR/Cas9 KO Plasmid (h) is a pool of plasmids designed for targeted disruption of the HCRT gene in human cell lines. Each plasmid co-expresses a unique single guide RNA (sgRNA) targeting a distinct site within the HCRT 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 HCRT 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 Orexin-A protein expression.
This CRISPR knockout system enables efficient generation of HCRT-deficient cell models for investigation of Orexin-A signaling, functional genomics studies, cancer biology research, and evaluation of therapeutic responses in human cell lines.
Key Features
- sgRNAs targeting HCRT exon(s) critical for Orexin-A 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 HCRT genomic sites to improve knockout efficiency
- Compatible with delivery by transfection
Design Variants
CRISPRs +/- HDRs
- gRNAs encoded by Orexin-A CRISPR/Cas9 KO Plasmid (h) and Orexin-A CRISPR/Cas9 KO Plasmid (h2) target distinct sites within the HCRT locus. One or both targeting designs may be available. See Related Products for availability.
- HDR donor constructs encoded by Orexin-A HDR Plasmid (h) and Orexin-A HDR Plasmid (h2) contain a puromycin resistance cassette and an RFP reporter flanked by HCRT homology arms to support homology-directed repair at defined HCRT 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.