Midnolin CRISPR/Cas9 KO Plasmid (m): sc-425598
- Target species: mouse
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- Midnolin 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 Midnolin 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
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
Midnolin CRISPR/Cas9 KO Plasmid (m) | sc-425598 | 20 µg | $397.00 |
Overview
Midn (midnolin) encodes a nucleolar-associated protein implicated in the regulation of transcriptional programs and proteostasis, with reported links to neuronal differentiation and cellular stress responses. Midnolin is thought to participate in nuclear processes that influence ribosome biogenesis and protein quality control, consistent with its enrichment in nucleolar and nuclear compartments. Altered MIDN expression has been associated with neurobiological phenotypes and has been explored in the context of dopaminergic neuron function and neurodegeneration-relevant pathways. In mouse models, Midn provides a tractable node for dissecting how nucleolar signaling interfaces with gene regulation in development and stress adaptation.
Midnolin CRISPR/Cas9 KO Plasmid (m) is a pool of plasmids designed for targeted disruption of the Midn gene in mouse cell lines. Each plasmid co-expresses a unique single guide RNA (sgRNA) targeting a distinct site within the Midn 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 Midn 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 Midnolin protein expression.
This CRISPR knockout system enables efficient generation of Midn-deficient cell models for investigation of Midnolin signaling, functional genomics studies, cancer biology research, and evaluation of therapeutic responses in human cell lines.
Key Features
- sgRNAs targeting Midn exon(s) critical for Midnolin 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 Midn genomic sites to improve knockout efficiency
- Compatible with delivery by transfection
Design Variants
CRISPRs +/- HDRs
- gRNAs encoded by Midnolin CRISPR/Cas9 KO Plasmid (m) and Midnolin CRISPR/Cas9 KO Plasmid (m2) target distinct sites within the Midn locus. One or both targeting designs may be available. See Related Products for availability.
- HDR donor constructs encoded by Midnolin HDR Plasmid (m) and Midnolin HDR Plasmid (m2) contain a puromycin resistance cassette and an RFP reporter flanked by Midn homology arms to support homology-directed repair at defined Midn 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.