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.
FLG/Filaggrin CRISPR/Cas9 KO Plasmid (h): sc-400644
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- FLG/Filaggrin 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 FLG/Filaggrin 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: FLG/Filaggrin Antibody (AKH1): sc-66192
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
FLG/Filaggrin CRISPR/Cas9 KO Plasmid (h) | sc-400644 | 20 µg | $397.00 |
Overview
FLG encodes filaggrin, a key epidermal differentiation protein produced as profilaggrin in keratohyalin granules and processed into filaggrin repeats that aggregate keratin intermediate filaments during terminal keratinocyte maturation. Filaggrin breakdown products contribute to natural moisturizing factor, supporting stratum corneum hydration, pH homeostasis, and barrier integrity. Through these functions, FLG intersects with cornified envelope formation, lipid organization, and inflammatory signaling linked to barrier stress. Loss-of-function variation or reduced expression is associated with impaired skin barrier phenotypes and is widely studied in atopic dermatitis and ichthyosis vulgaris models.
FLG/Filaggrin CRISPR/Cas9 KO Plasmid (h) is a pool of plasmids designed for targeted disruption of the FLG gene in human cell lines. Each plasmid co-expresses a unique single guide RNA (sgRNA) targeting a distinct site within the FLG 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 FLG 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 FLG/Filaggrin protein expression.
This CRISPR knockout system enables efficient generation of FLG-deficient cell models for investigation of FLG/Filaggrin signaling, functional genomics studies, cancer biology research, and evaluation of therapeutic responses in human cell lines.
Key Features
- sgRNAs targeting FLG exon(s) critical for FLG/Filaggrin 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 FLG genomic sites to improve knockout efficiency
- Compatible with delivery by transfection
Design Variants
CRISPRs +/- HDRs
- gRNAs encoded by FLG/Filaggrin CRISPR/Cas9 KO Plasmid (h) and FLG/Filaggrin CRISPR/Cas9 KO Plasmid (h2) target distinct sites within the FLG locus. One or both targeting designs may be available. See Related Products for availability.
- HDR donor constructs encoded by FLG/Filaggrin HDR Plasmid (h) and FLG/Filaggrin HDR Plasmid (h2) contain a puromycin resistance cassette and an RFP reporter flanked by FLG homology arms to support homology-directed repair at defined FLG 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.