POLR3B CRISPR/Cas9 KO Plasmid (h): sc-406739
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- POLR3B 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 POLR3B 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: POLR3B Antibody (G-6): sc-515362
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
POLR3B CRISPR/Cas9 KO Plasmid (h) | sc-406739 | 20 µg | $397.00 |
Overview
POLR3B encodes the second-largest subunit of RNA polymerase III, a multi-subunit enzyme responsible for transcription of small noncoding RNAs including tRNAs, 5S rRNA, and other short RNAs required for translation and cellular growth. Through regulation of Pol III output, POLR3B influences ribosome biogenesis, proteostasis, and metabolic adaptation, integrating nutrient- and stress-responsive signaling with biosynthetic capacity. Perturbation of Pol III transcription impacts innate immune sensing of cytosolic nucleic acids and broader transcriptional homeostasis in proliferative and differentiated cell states. Pathogenic variants in POLR3B are linked to Pol III–related leukodystrophy spectrum disorders and neurodevelopmental phenotypes, making it a key gene for studying transcriptional machinery dysfunction in human cells.
POLR3B CRISPR/Cas9 KO Plasmid (h) is a pool of plasmids designed for targeted disruption of the POLR3B gene in human cell lines. Each plasmid co-expresses a unique single guide RNA (sgRNA) targeting a distinct site within the POLR3B 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 POLR3B 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 POLR3B protein expression.
This CRISPR knockout system enables efficient generation of POLR3B-deficient cell models for investigation of POLR3B signaling, functional genomics studies, cancer biology research, and evaluation of therapeutic responses in human cell lines.
Key Features
- sgRNAs targeting POLR3B exon(s) critical for POLR3B 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 POLR3B genomic sites to improve knockout efficiency
- Compatible with delivery by transfection
Design Variants
CRISPRs +/- HDRs
- gRNAs encoded by POLR3B CRISPR/Cas9 KO Plasmid (h) and POLR3B CRISPR/Cas9 KO Plasmid (h2) target distinct sites within the POLR3B locus. One or both targeting designs may be available. See Related Products for availability.
- HDR donor constructs encoded by POLR3B HDR Plasmid (h) and POLR3B HDR Plasmid (h2) contain a puromycin resistance cassette and an RFP reporter flanked by POLR3B homology arms to support homology-directed repair at defined POLR3B 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.