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.
RPA40 CRISPR/Cas9 KO Plasmid (h): sc-404594
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- RPA40 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 RPA40 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: RPA40 Antibody (H-6): sc-374443
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
RPA40 CRISPR/Cas9 KO Plasmid (h) | sc-404594 | 20 µg | $397.00 |
Overview
POLR1C encodes RPA40, a shared subunit of RNA polymerase I and RNA polymerase III that supports assembly and stability of these transcription complexes. Through its roles in rRNA synthesis by Pol I and transcription of tRNAs and other small noncoding RNAs by Pol III, RPA40 contributes to ribosome biogenesis, proteostasis, and cell growth control. Disruption of POLR1C-linked transcriptional output can alter nucleolar function and cellular stress responses, connecting this factor to pathways that influence proliferation and differentiation. Variants in POLR1C have been associated with inherited disorders affecting myelin and craniofacial development, underscoring the importance of Pol I/III integrity in human biology.
RPA40 CRISPR/Cas9 KO Plasmid (h) is a pool of plasmids designed for targeted disruption of the POLR1C gene in human cell lines. Each plasmid co-expresses a unique single guide RNA (sgRNA) targeting a distinct site within the POLR1C 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 POLR1C 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 RPA40 protein expression.
This CRISPR knockout system enables efficient generation of POLR1C-deficient cell models for investigation of RPA40 signaling, functional genomics studies, cancer biology research, and evaluation of therapeutic responses in human cell lines.
Key Features
- sgRNAs targeting POLR1C exon(s) critical for RPA40 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 POLR1C genomic sites to improve knockout efficiency
- Compatible with delivery by transfection
Design Variants
CRISPRs +/- HDRs
- gRNAs encoded by RPA40 CRISPR/Cas9 KO Plasmid (h) and RPA40 CRISPR/Cas9 KO Plasmid (h2) target distinct sites within the POLR1C locus. One or both targeting designs may be available. See Related Products for availability.
- HDR donor constructs encoded by RPA40 HDR Plasmid (h) and RPA40 HDR Plasmid (h2) contain a puromycin resistance cassette and an RFP reporter flanked by POLR1C homology arms to support homology-directed repair at defined POLR1C 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.