RPA135 CRISPR/Cas9 KO Plasmid (h): sc-402818
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- RPA135 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 RPA135 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: RPA135 Antibody (4H6): sc-293272
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
RPA135 CRISPR/Cas9 KO Plasmid (h) | sc-402818 | 20 µg | $397.00 |
Overview
POLR1B encodes RPA135, the second-largest catalytic subunit of RNA polymerase I that drives 47S pre-rRNA synthesis in the nucleolus. RPA135 is central to ribosome biogenesis and couples growth-factor signaling and nutrient status to rDNA transcription, linking it to nucleolar organization, cell-cycle progression, and proteostasis. Perturbation of Pol I transcription activates nucleolar stress pathways that converge on p53-dependent checkpoints and can reshape global translation programs. Dysregulated rRNA production and altered ribosome biogenesis are recurrent features of proliferative and stress-adaptation states, making POLR1B a useful node for studying transcriptional control of rDNA and nucleolar function.
RPA135 CRISPR/Cas9 KO Plasmid (h) is a pool of plasmids designed for targeted disruption of the POLR1B gene in human cell lines. Each plasmid co-expresses a unique single guide RNA (sgRNA) targeting a distinct site within the POLR1B 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 POLR1B 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 RPA135 protein expression.
This CRISPR knockout system enables efficient generation of POLR1B-deficient cell models for investigation of RPA135 signaling, functional genomics studies, cancer biology research, and evaluation of therapeutic responses in human cell lines.
Key Features
- sgRNAs targeting POLR1B exon(s) critical for RPA135 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 POLR1B genomic sites to improve knockout efficiency
- Compatible with delivery by transfection
Design Variants
CRISPRs +/- HDRs
- gRNAs encoded by RPA135 CRISPR/Cas9 KO Plasmid (h) and RPA135 CRISPR/Cas9 KO Plasmid (h2) target distinct sites within the POLR1B locus. One or both targeting designs may be available. See Related Products for availability.
- HDR donor constructs encoded by RPA135 HDR Plasmid (h) and RPA135 HDR Plasmid (h2) contain a puromycin resistance cassette and an RFP reporter flanked by POLR1B homology arms to support homology-directed repair at defined POLR1B 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.