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.
U2AF65 CRISPR/Cas9 KO Plasmid (m): sc-423572
- Target species: mouse
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- U2AF65 CRISPR/Cas9 Knockout (KO) Plasmid (m) 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 U2AF65 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: U2AF65 Antibody (MC3): sc-53942
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
U2AF65 CRISPR/Cas9 KO Plasmid (m) | sc-423572 | 20 µg | $397.00 |
Overview
U2af2 encodes U2AF65, an essential RNA-binding component of the U2 auxiliary factor that recognizes the polypyrimidine tract at 3′ splice sites and cooperates with U2AF35 to recruit the U2 snRNP during early spliceosome assembly. By coupling splice-site selection to exon definition, U2AF65 helps shape global alternative splicing programs that regulate cell-cycle progression, differentiation, and stress responses. Perturbation of U2AF65-dependent splicing can alter isoform balance in pathways controlling DNA damage responses and apoptosis, making U2af2 a useful node for dissecting RNA processing–driven phenotypes. Dysregulated splice-site recognition and altered U2AF complex activity are broadly relevant to mechanisms underlying developmental abnormalities and cancer-associated transcriptome remodeling.
U2AF65 CRISPR/Cas9 KO Plasmid (m) is a pool of plasmids designed for targeted disruption of the U2af2 gene in mouse cell lines. Each plasmid co-expresses a unique single guide RNA (sgRNA) targeting a distinct site within the U2af2 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 U2af2 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 U2AF65 protein expression.
This CRISPR knockout system enables efficient generation of U2af2-deficient cell models for investigation of U2AF65 signaling, functional genomics studies, cancer biology research, and evaluation of therapeutic responses in human cell lines.
Key Features
- sgRNAs targeting U2af2 exon(s) critical for U2AF65 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 U2af2 genomic sites to improve knockout efficiency
- Compatible with delivery by transfection
Design Variants
CRISPRs +/- HDRs
- gRNAs encoded by U2AF65 CRISPR/Cas9 KO Plasmid (m) and U2AF65 CRISPR/Cas9 KO Plasmid (m2) target distinct sites within the U2af2 locus. One or both targeting designs may be available. See Related Products for availability.
- HDR donor constructs encoded by U2AF65 HDR Plasmid (m) and U2AF65 HDR Plasmid (m2) contain a puromycin resistance cassette and an RFP reporter flanked by U2af2 homology arms to support homology-directed repair at defined U2af2 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.