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.
NF-L CRISPR/Cas9 KO Plasmid (h): sc-402254
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- NF-L 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 NF-L 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: NF-L Antibody (8A1): sc-20012
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
NF-L CRISPR/Cas9 KO Plasmid (h) | sc-402254 | 20 µg | $397.00 |
Overview
NEFL encodes neurofilament light chain (NF-L), a core intermediate filament protein that assembles with NEFM and NEFH to form the neuronal cytoskeleton and maintain axonal caliber and mechanical stability. NF-L participates in neurofilament transport and organization, influencing axonal trafficking, cytoskeletal dynamics, and long-range neuronal connectivity. Disrupted NEFL function perturbs neurofilament assembly and axonal integrity, linking cytoskeletal stress pathways to progressive neuronal dysfunction. Variants and altered NF-L homeostasis are associated with inherited neuropathies and neurodegenerative processes where axonal structure and transport are compromised.
NF-L CRISPR/Cas9 KO Plasmid (h) is a pool of plasmids designed for targeted disruption of the NEFL gene in human cell lines. Each plasmid co-expresses a unique single guide RNA (sgRNA) targeting a distinct site within the NEFL 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 NEFL 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 NF-L protein expression.
This CRISPR knockout system enables efficient generation of NEFL-deficient cell models for investigation of NF-L signaling, functional genomics studies, cancer biology research, and evaluation of therapeutic responses in human cell lines.
Key Features
- sgRNAs targeting NEFL exon(s) critical for NF-L 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 NEFL genomic sites to improve knockout efficiency
- Compatible with delivery by transfection
Design Variants
CRISPRs +/- HDRs
- gRNAs encoded by NF-L CRISPR/Cas9 KO Plasmid (h) and NF-L CRISPR/Cas9 KO Plasmid (h2) target distinct sites within the NEFL locus. One or both targeting designs may be available. See Related Products for availability.
- HDR donor constructs encoded by NF-L HDR Plasmid (h) and NF-L HDR Plasmid (h2) contain a puromycin resistance cassette and an RFP reporter flanked by NEFL homology arms to support homology-directed repair at defined NEFL 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.