Nebulette CRISPR/Cas9 KO Plasmid (h): sc-406565
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- Nebulette 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 Nebulette 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: Nebulette Antibody (G-9): sc-393784
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
Nebulette CRISPR/Cas9 KO Plasmid (h) | sc-406565 | 20 µg | $397.00 |
Overview
NEBL encodes nebulette, a cardiac-specific actin-binding protein of the Z-disc that helps organize thin filaments and stabilize sarcomere architecture in striated muscle. Nebulette links actin filaments to Z-line scaffolds and contributes to mechanotransduction and force transmission during myofibril assembly and contraction. Through interactions with cytoskeletal and Z-disc proteins, NEBL supports cardiomyocyte structural integrity and stress-responsive remodeling pathways. Genetic variation or dysregulated expression of NEBL has been associated with cardiomyopathies and related cardiac muscle disorders, making it relevant for studies of sarcomere maintenance and disease-linked cytoskeletal disruption.
Nebulette CRISPR/Cas9 KO Plasmid (h) is a pool of plasmids designed for targeted disruption of the NEBL gene in human cell lines. Each plasmid co-expresses a unique single guide RNA (sgRNA) targeting a distinct site within the NEBL 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 NEBL 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 Nebulette protein expression.
This CRISPR knockout system enables efficient generation of NEBL-deficient cell models for investigation of Nebulette signaling, functional genomics studies, cancer biology research, and evaluation of therapeutic responses in human cell lines.
Key Features
- sgRNAs targeting NEBL exon(s) critical for Nebulette 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 NEBL genomic sites to improve knockout efficiency
- Compatible with delivery by transfection
Design Variants
CRISPRs +/- HDRs
- gRNAs encoded by Nebulette CRISPR/Cas9 KO Plasmid (h) and Nebulette CRISPR/Cas9 KO Plasmid (h2) target distinct sites within the NEBL locus. One or both targeting designs may be available. See Related Products for availability.
- HDR donor constructs encoded by Nebulette HDR Plasmid (h) and Nebulette HDR Plasmid (h2) contain a puromycin resistance cassette and an RFP reporter flanked by NEBL homology arms to support homology-directed repair at defined NEBL 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.