ANO1 CRISPR/Cas9 KO Plasmid (h2): sc-401314-KO-2
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- ANO1 CRISPR/Cas9 Knockout (KO) Plasmid (h2) 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 ANO1 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: ANO1 Antibody (C-5): sc-377115
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
ANO1 CRISPR/Cas9 KO Plasmid (h2) | sc-401314-KO-2 | 20 µg | $397.00 |
Overview
ANO1 (TMEM16A) encodes a Ca2+-activated chloride channel that regulates epithelial ion and fluid transport, smooth muscle excitability, and sensory signal transduction. Channel activity couples intracellular Ca2+ signals to membrane depolarization and chloride flux, shaping processes such as airway and gastrointestinal secretion and modulation of neuronal firing. ANO1 participates in signaling networks downstream of receptor-driven phospholipase C activation and Ca2+ release, and it influences cell motility and membrane dynamics in multiple tissue contexts. Dysregulated ANO1 expression or activity has been associated with altered excitability and secretion phenotypes and has been frequently studied in oncology and inflammatory settings as a contributor to aberrant cellular proliferation and migration programs.
ANO1 CRISPR/Cas9 KO Plasmid (h2) is a pool of plasmids designed for targeted disruption of the ANO1 gene in human cell lines. Each plasmid co-expresses a unique single guide RNA (sgRNA) targeting a distinct site within the ANO1 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 ANO1 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 ANO1 protein expression.
This CRISPR knockout system enables efficient generation of ANO1-deficient cell models for investigation of ANO1 signaling, functional genomics studies, cancer biology research, and evaluation of therapeutic responses in human cell lines.
Key Features
- sgRNAs targeting ANO1 exon(s) critical for ANO1 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 ANO1 genomic sites to improve knockout efficiency
- Compatible with delivery by transfection
Design Variants
CRISPRs +/- HDRs
- gRNAs encoded by ANO1 CRISPR/Cas9 KO Plasmid (h) and ANO1 CRISPR/Cas9 KO Plasmid (h2) target distinct sites within the ANO1 locus. One or both targeting designs may be available. See Related Products for availability.
- HDR donor constructs encoded by ANO1 HDR Plasmid (h) and ANO1 HDR Plasmid (h2) contain a puromycin resistance cassette and an RFP reporter flanked by ANO1 homology arms to support homology-directed repair at defined ANO1 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.