PAP-β CRISPR/Cas9 KO Plasmid (h): sc-403907
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- PAP-β 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 PAP-β 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
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
PAP-β CRISPR/Cas9 KO Plasmid (h) | sc-403907 | 20 µg | $397.00 |
Overview
PAPOLB encodes poly(A) polymerase beta (PAP-β), a non-canonical polyadenylation enzyme with enriched expression in male germ cells that supports post-transcriptional control during spermatogenesis. By extending poly(A) tails on select mRNAs, PAP-β influences transcript stability, nuclear export, and translational activation, integrating with broader RNA processing networks and mRNA surveillance. Altered PAPOLB activity has been linked to defects in sperm development and impaired fertility phenotypes, making it relevant for studies of germline gene regulation. PAPOLB also provides a model for dissecting how specialized poly(A) polymerases shape stage-specific gene expression programs.
PAP-β CRISPR/Cas9 KO Plasmid (h) is a pool of plasmids designed for targeted disruption of the PAPOLB gene in human cell lines. Each plasmid co-expresses a unique single guide RNA (sgRNA) targeting a distinct site within the PAPOLB 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 PAPOLB 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 PAP-β protein expression.
This CRISPR knockout system enables efficient generation of PAPOLB-deficient cell models for investigation of PAP-β signaling, functional genomics studies, cancer biology research, and evaluation of therapeutic responses in human cell lines.
Key Features
- sgRNAs targeting PAPOLB exon(s) critical for PAP-β 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 PAPOLB genomic sites to improve knockout efficiency
- Compatible with delivery by transfection
Design Variants
CRISPRs +/- HDRs
- gRNAs encoded by PAP-β CRISPR/Cas9 KO Plasmid (h) and PAP-β CRISPR/Cas9 KO Plasmid (h2) target distinct sites within the PAPOLB locus. One or both targeting designs may be available. See Related Products for availability.
- HDR donor constructs encoded by PAP-β HDR Plasmid (h) and PAP-β HDR Plasmid (h2) contain a puromycin resistance cassette and an RFP reporter flanked by PAPOLB homology arms to support homology-directed repair at defined PAPOLB 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.