betaKlotho CRISPR/Cas9 KO Plasmid (h): sc-401614
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- betaKlotho 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 betaKlotho 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
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
betaKlotho CRISPR/Cas9 KO Plasmid (h) | sc-401614 | 20 µg | $397.00 |
Overview
KLB encodes betaKlotho, a single-pass transmembrane co-receptor that confers ligand specificity to endocrine fibroblast growth factors, most prominently FGF19 and FGF21, through complex formation with FGFRs. In human tissues such as liver and adipose, betaKlotho-dependent signaling regulates MAPK/ERK pathway activation and transcriptional programs controlling bile acid homeostasis, lipid utilization, glucose metabolism, and energy balance. Altered KLB expression or betaKlotho function has been linked to metabolic dysregulation, including obesity-associated phenotypes, insulin resistance, and nonalcoholic fatty liver disease, and can modulate stress and inflammatory signaling in metabolically active cells. These features make KLB a useful node for dissecting endocrine FGF pathways and tissue-specific metabolic crosstalk.
betaKlotho CRISPR/Cas9 KO Plasmid (h) is a pool of plasmids designed for targeted disruption of the KLB gene in human cell lines. Each plasmid co-expresses a unique single guide RNA (sgRNA) targeting a distinct site within the KLB 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 KLB 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 betaKlotho protein expression.
This CRISPR knockout system enables efficient generation of KLB-deficient cell models for investigation of betaKlotho signaling, functional genomics studies, cancer biology research, and evaluation of therapeutic responses in human cell lines.
Key Features
- sgRNAs targeting KLB exon(s) critical for betaKlotho 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 KLB genomic sites to improve knockout efficiency
- Compatible with delivery by transfection
Design Variants
CRISPRs +/- HDRs
- gRNAs encoded by betaKlotho CRISPR/Cas9 KO Plasmid (h) and betaKlotho CRISPR/Cas9 KO Plasmid (h2) target distinct sites within the KLB locus. One or both targeting designs may be available. See Related Products for availability.
- HDR donor constructs encoded by betaKlotho HDR Plasmid (h) and betaKlotho HDR Plasmid (h2) contain a puromycin resistance cassette and an RFP reporter flanked by KLB homology arms to support homology-directed repair at defined KLB 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.