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.
TFIIIB90-1 CRISPR/Cas9 KO Plasmid (h): sc-405826
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- TFIIIB90-1 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 TFIIIB90-1 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: TFIIIB90-1/2/3/5 Antibody (A-8): sc-390821
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
TFIIIB90-1 CRISPR/Cas9 KO Plasmid (h) | sc-405826 | 20 µg | $397.00 |
Overview
BRF1 encodes the TFIIIB90-1 subunit of transcription factor IIIB, a core component of the RNA polymerase III initiation machinery required for transcription of tRNAs, 5S rRNA, and other small noncoding RNAs. Through assembly with TBP and BDP1 at Pol III promoters, TFIIIB90-1 supports chromatin engagement, preinitiation complex formation, and regulation of cellular translation capacity and growth programs. Pol III output is tightly coupled to nutrient sensing and proliferative signaling, linking BRF1 function to ribosome biogenesis, proteostasis, and stress adaptation. Dysregulated Pol III transcription and altered TFIIIB activity are frequently associated with oncogenic states and other disorders characterized by aberrant biosynthetic demand and cell-cycle control.
TFIIIB90-1 CRISPR/Cas9 KO Plasmid (h) is a pool of plasmids designed for targeted disruption of the BRF1 gene in human cell lines. Each plasmid co-expresses a unique single guide RNA (sgRNA) targeting a distinct site within the BRF1 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 BRF1 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 TFIIIB90-1 protein expression.
This CRISPR knockout system enables efficient generation of BRF1-deficient cell models for investigation of TFIIIB90-1 signaling, functional genomics studies, cancer biology research, and evaluation of therapeutic responses in human cell lines.
Key Features
- sgRNAs targeting BRF1 exon(s) critical for TFIIIB90-1 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 BRF1 genomic sites to improve knockout efficiency
- Compatible with delivery by transfection
Design Variants
CRISPRs +/- HDRs
- gRNAs encoded by TFIIIB90-1 CRISPR/Cas9 KO Plasmid (h) and TFIIIB90-1 CRISPR/Cas9 KO Plasmid (h2) target distinct sites within the BRF1 locus. One or both targeting designs may be available. See Related Products for availability.
- HDR donor constructs encoded by TFIIIB90-1 HDR Plasmid (h) and TFIIIB90-1 HDR Plasmid (h2) contain a puromycin resistance cassette and an RFP reporter flanked by BRF1 homology arms to support homology-directed repair at defined BRF1 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.