Not CRISPR data: the image illustrates antibody quality for visualizing CRISPR experiment results.
Not CRISPR data: the image illustrates antibody quality for visualizing CRISPR experiment results.
nucleobindin CRISPR/Cas9 KO Plasmid (h): sc-405167
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- nucleobindin 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 nucleobindin 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: nucleobindin Antibody (D-8): sc-515246
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
nucleobindin CRISPR/Cas9 KO Plasmid (h) | sc-405167 | 20 µg | $397.00 |
Overview
NUCB1 encodes nucleobindin, a calcium-binding protein that localizes to the secretory pathway and participates in Ca2+-dependent regulation of intracellular trafficking and protein sorting. Through EF-hand motifs and interactions with luminal cargo, NUCB1 has been linked to Golgi/ER homeostasis, vesicle-mediated transport, and stress-adaptive secretory responses. Altered NUCB1 expression has been reported across contexts involving dysregulated proteostasis and cellular stress signaling, making it relevant for mechanistic studies of secretion, membrane dynamics, and Ca2+-regulated pathways. NUCB1-dependent processes are also examined in disease models where secretion, aggregation, or organelle stress responses contribute to pathophysiology.
nucleobindin CRISPR/Cas9 KO Plasmid (h) is a pool of plasmids designed for targeted disruption of the NUCB1 gene in human cell lines. Each plasmid co-expresses a unique single guide RNA (sgRNA) targeting a distinct site within the NUCB1 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 NUCB1 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 nucleobindin protein expression.
This CRISPR knockout system enables efficient generation of NUCB1-deficient cell models for investigation of nucleobindin signaling, functional genomics studies, cancer biology research, and evaluation of therapeutic responses in human cell lines.
Key Features
- sgRNAs targeting NUCB1 exon(s) critical for nucleobindin 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 NUCB1 genomic sites to improve knockout efficiency
- Compatible with delivery by transfection
Design Variants
CRISPRs +/- HDRs
- gRNAs encoded by nucleobindin CRISPR/Cas9 KO Plasmid (h) and nucleobindin CRISPR/Cas9 KO Plasmid (h2) target distinct sites within the NUCB1 locus. One or both targeting designs may be available. See Related Products for availability.
- HDR donor constructs encoded by nucleobindin HDR Plasmid (h) and nucleobindin HDR Plasmid (h2) contain a puromycin resistance cassette and an RFP reporter flanked by NUCB1 homology arms to support homology-directed repair at defined NUCB1 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.