POL H CRISPR/Cas9 KO Plasmid (h2): sc-401794-KO-2

Overview

POLH encodes DNA polymerase eta (Pol η), a Y-family translesion synthesis polymerase that enables accurate bypass of UV-induced cyclobutane pyrimidine dimers during DNA replication. It functions within DNA damage tolerance and replication stress responses, coordinating with proliferating cell nuclear antigen (PCNA) and ubiquitin-dependent switching from replicative polymerases to specialized TLS polymerases. Loss or dysfunction of Pol η compromises genome stability and elevates UV mutagenesis, linking POLH to xeroderma pigmentosum variant (XP-V) and broader mechanisms of carcinogenesis driven by DNA repair defects. Because POLH sits at the intersection of nucleotide excision repair–associated lesion processing and S-phase checkpoint adaptation, it is frequently studied in pathways governing mutational signatures and replication fork dynamics.

POL H CRISPR/Cas9 KO Plasmid (h2) is a pool of plasmids designed for targeted disruption of the POLH gene in human cell lines. Each plasmid co-expresses a unique single guide RNA (sgRNA) targeting a distinct site within the POLH 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 POLH 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 H protein expression.

This CRISPR knockout system enables efficient generation of POLH-deficient cell models for investigation of POL H signaling, functional genomics studies, cancer biology research, and evaluation of therapeutic responses in human cell lines.

Key Features

• sgRNAs targeting POLH exon(s) critical for POL H 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 POLH genomic sites to improve knockout efficiency
• Compatible with delivery by transfection

Design Variants

CRISPRs +/- HDRs

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