Pol I/II/III RPB8 CRISPR/Cas9 KO Plasmid (h): sc-406520

Overview

POLR2H encodes RPB8, a small, conserved subunit shared by RNA polymerases I, II, and III that contributes to polymerase assembly, stability, and transcriptional competence. Through its participation in rRNA synthesis (Pol I), mRNA transcription (Pol II), and tRNA/5S rRNA production (Pol III), RPB8 supports core gene expression programs that couple transcription with ribosome biogenesis, cell-cycle control, and proteostasis. Altered activity of basal transcription machinery and nucleolar stress pathways can reshape global transcript output and downstream signaling, making POLR2H a useful node for studying transcriptional vulnerabilities and genome-wide expression rewiring in disease-relevant contexts. Its essential, multi-polymerase role also provides a mechanistic handle to interrogate how general transcription factors and chromatin states coordinate across polymerase systems.

Pol I/II/III RPB8 CRISPR/Cas9 KO Plasmid (h) is a pool of plasmids designed for targeted disruption of the POLR2H gene in human cell lines. Each plasmid co-expresses a unique single guide RNA (sgRNA) targeting a distinct site within the POLR2H 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 POLR2H 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 Pol I/II/III RPB8 protein expression.

This CRISPR knockout system enables efficient generation of POLR2H-deficient cell models for investigation of Pol I/II/III RPB8 signaling, functional genomics studies, cancer biology research, and evaluation of therapeutic responses in human cell lines.

Key Features

• sgRNAs targeting POLR2H exon(s) critical for Pol I/II/III RPB8 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 POLR2H genomic sites to improve knockout efficiency
• Compatible with delivery by transfection

Design Variants

CRISPRs +/- HDRs

• gRNAs encoded by Pol I/II/III RPB8 CRISPR/Cas9 KO Plasmid (h) and Pol I/II/III RPB8 CRISPR/Cas9 KO Plasmid (h2) target distinct sites within the POLR2H locus. One or both targeting designs may be available. See Related Products for availability.
• HDR donor constructs encoded by Pol I/II/III RPB8 HDR Plasmid (h) and Pol I/II/III RPB8 HDR Plasmid (h2) contain a puromycin resistance cassette and an RFP reporter flanked by POLR2H homology arms to support homology-directed repair at defined POLR2H 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.