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.
Bob 1 CRISPR/Cas9 KO Plasmid (h): sc-400771
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- Bob 1 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 Bob 1 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: Bob 1 Antibody (6F10): sc-23932
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
Bob 1 CRISPR/Cas9 KO Plasmid (h) | sc-400771 | 20 µg | $397.00 |
Overview
POU2AF1 (Bob 1/OBF-1) is a B cell–restricted transcriptional coactivator that binds OCT1/POU2F1 and OCT2/POU2F2 to potentiate immunoglobulin gene transcription and broader B cell gene expression programs. It supports germinal center responses, class-switch recombination, and plasma cell differentiation by integrating enhancer activity at octamer motifs with signaling inputs that shape B cell activation and survival. Dysregulated POU2AF1 expression and associated transcriptional circuitry have been reported in B cell malignancies, including lymphomas and leukemias, and in immune phenotypes linked to altered humoral responses. As such, Bob 1 is commonly studied in pathways controlling B cell identity, antigen-driven maturation, and transcriptional regulation of immune effector genes.
Bob 1 CRISPR/Cas9 KO Plasmid (h) is a pool of plasmids designed for targeted disruption of the POU2AF1 gene in human cell lines. Each plasmid co-expresses a unique single guide RNA (sgRNA) targeting a distinct site within the POU2AF1 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 POU2AF1 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 Bob 1 protein expression.
This CRISPR knockout system enables efficient generation of POU2AF1-deficient cell models for investigation of Bob 1 signaling, functional genomics studies, cancer biology research, and evaluation of therapeutic responses in human cell lines.
Key Features
- sgRNAs targeting POU2AF1 exon(s) critical for Bob 1 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 POU2AF1 genomic sites to improve knockout efficiency
- Compatible with delivery by transfection
Design Variants
CRISPRs +/- HDRs
- gRNAs encoded by Bob 1 CRISPR/Cas9 KO Plasmid (h) and Bob 1 CRISPR/Cas9 KO Plasmid (h2) target distinct sites within the POU2AF1 locus. One or both targeting designs may be available. See Related Products for availability.
- HDR donor constructs encoded by Bob 1 HDR Plasmid (h) and Bob 1 HDR Plasmid (h2) contain a puromycin resistance cassette and an RFP reporter flanked by POU2AF1 homology arms to support homology-directed repair at defined POU2AF1 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.