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.
AFF4 CRISPR/Cas9 KO Plasmid (h): sc-406238
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- AFF4 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 AFF4 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: AFF4 Antibody (G-1): sc-390310
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
AFF4 CRISPR/Cas9 KO Plasmid (h) | sc-406238 | 20 µg | $397.00 |
Overview
AFF4 (AF4/FMR2 family member 4) encodes a nuclear transcriptional regulator that functions as a core component of the Super Elongation Complex (SEC), coordinating the transition of RNA polymerase II into productive elongation. Through interactions with elongation factors such as P-TEFb and ELL proteins, AFF4 supports rapid induction of stimulus-responsive genes and influences chromatin-associated transcriptional programs. AFF4-dependent control of transcriptional elongation contributes to regulation of cell identity, proliferation, and stress-response pathways. Dysregulated SEC activity and altered AFF4-associated transcriptional circuitry are relevant to oncogenic gene expression programs and other diseases driven by aberrant RNA polymerase II elongation.
AFF4 CRISPR/Cas9 KO Plasmid (h) is a pool of plasmids designed for targeted disruption of the AFF4 gene in human cell lines. Each plasmid co-expresses a unique single guide RNA (sgRNA) targeting a distinct site within the AFF4 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 AFF4 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 AFF4 protein expression.
This CRISPR knockout system enables efficient generation of AFF4-deficient cell models for investigation of AFF4 signaling, functional genomics studies, cancer biology research, and evaluation of therapeutic responses in human cell lines.
Key Features
- sgRNAs targeting AFF4 exon(s) critical for AFF4 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 AFF4 genomic sites to improve knockout efficiency
- Compatible with delivery by transfection
Design Variants
CRISPRs +/- HDRs
- gRNAs encoded by AFF4 CRISPR/Cas9 KO Plasmid (h) and AFF4 CRISPR/Cas9 KO Plasmid (h2) target distinct sites within the AFF4 locus. One or both targeting designs may be available. See Related Products for availability.
- HDR donor constructs encoded by AFF4 HDR Plasmid (h) and AFF4 HDR Plasmid (h2) contain a puromycin resistance cassette and an RFP reporter flanked by AFF4 homology arms to support homology-directed repair at defined AFF4 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.