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.
THP CRISPR/Cas9 KO Plasmid (h): sc-401238
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- THP 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 THP 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: THP Antibody (B-2): sc-271022
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
THP CRISPR/Cas9 KO Plasmid (h) | sc-401238 | 20 µg | $397.00 |
Overview
UMOD encodes uromodulin (THP), a highly abundant glycoprotein produced by epithelial cells of the thick ascending limb and secreted into urine following processing in the secretory pathway. THP contributes to tubular homeostasis through polymer formation, regulation of ion transport processes, and interactions with immune and extracellular matrix components in the urinary tract. Cellular phenotypes linked to UMOD include altered protein folding/trafficking, ER stress, and disrupted tubule function, reflecting its sensitivity to maturation and apical release. Genetic variation or dysfunction in UMOD is associated with inherited kidney disorders and susceptibility to chronic kidney disease, making it a useful locus for studying renal epithelial biology and proteostasis pathways.
THP CRISPR/Cas9 KO Plasmid (h) is a pool of plasmids designed for targeted disruption of the UMOD gene in human cell lines. Each plasmid co-expresses a unique single guide RNA (sgRNA) targeting a distinct site within the UMOD 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 UMOD 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 THP protein expression.
This CRISPR knockout system enables efficient generation of UMOD-deficient cell models for investigation of THP signaling, functional genomics studies, cancer biology research, and evaluation of therapeutic responses in human cell lines.
Key Features
- sgRNAs targeting UMOD exon(s) critical for THP 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 UMOD genomic sites to improve knockout efficiency
- Compatible with delivery by transfection
Design Variants
CRISPRs +/- HDRs
- gRNAs encoded by THP CRISPR/Cas9 KO Plasmid (h) and THP CRISPR/Cas9 KO Plasmid (h2) target distinct sites within the UMOD locus. One or both targeting designs may be available. See Related Products for availability.
- HDR donor constructs encoded by THP HDR Plasmid (h) and THP HDR Plasmid (h2) contain a puromycin resistance cassette and an RFP reporter flanked by UMOD homology arms to support homology-directed repair at defined UMOD 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.