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.
RNase H1 CRISPR/Cas9 KO Plasmid (m): sc-422686
- Target species: mouse
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- RNase H1 CRISPR/Cas9 Knockout (KO) Plasmid (m) 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 RNase H1 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: RNase H1 Antibody (H-4): sc-376326
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
RNase H1 CRISPR/Cas9 KO Plasmid (m) | sc-422686 | 20 µg | $397.00 |
Overview
Mouse Rnaseh1 encodes RNase H1, a structure-specific endonuclease that degrades the RNA strand of RNA:DNA hybrids and resolves R-loops generated during replication and transcription. By processing Okazaki fragment intermediates and limiting persistent R-loop formation, RNase H1 supports mitochondrial and nuclear genome stability, replication fork progression, and transcriptional homeostasis. Dysregulated RNA:DNA hybrid metabolism is linked to replication stress, DNA damage signaling, and mitochondrial dysfunction, making Rnaseh1 a useful node for studying genome maintenance pathways. Rnaseh1 perturbation is therefore relevant for mechanistic work in nucleic acid metabolism, mitochondrial biology, and stress-response phenotypes in mammalian cells.
RNase H1 CRISPR/Cas9 KO Plasmid (m) is a pool of plasmids designed for targeted disruption of the Rnaseh1 gene in mouse cell lines. Each plasmid co-expresses a unique single guide RNA (sgRNA) targeting a distinct site within the Rnaseh1 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 Rnaseh1 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 RNase H1 protein expression.
This CRISPR knockout system enables efficient generation of Rnaseh1-deficient cell models for investigation of RNase H1 signaling, functional genomics studies, cancer biology research, and evaluation of therapeutic responses in human cell lines.
Key Features
- sgRNAs targeting Rnaseh1 exon(s) critical for RNase H1 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 Rnaseh1 genomic sites to improve knockout efficiency
- Compatible with delivery by transfection
Design Variants
CRISPRs +/- HDRs
- gRNAs encoded by RNase H1 CRISPR/Cas9 KO Plasmid (m) and RNase H1 CRISPR/Cas9 KO Plasmid (m2) target distinct sites within the Rnaseh1 locus. One or both targeting designs may be available. See Related Products for availability.
- HDR donor constructs encoded by RNase H1 HDR Plasmid (m) and RNase H1 HDR Plasmid (m2) contain a puromycin resistance cassette and an RFP reporter flanked by Rnaseh1 homology arms to support homology-directed repair at defined Rnaseh1 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.