MGMT CRISPR/Cas9 KO Plasmid (m): sc-421647
- Target species: mouse
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- MGMT 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 MGMT 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: MGMT Antibody (E-1): sc-166528
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
MGMT CRISPR/Cas9 KO Plasmid (m) | sc-421647 | 20 µg | $397.00 |
Overview
Mouse Mgmt encodes O-6-methylguanine-DNA methyltransferase (MGMT), a direct DNA repair enzyme that removes alkyl adducts from the O6 position of guanine by transferring the lesion to an active-site cysteine in a single-use “suicide” reaction. This activity counteracts mutagenesis and replication stress arising from endogenous and environmental alkylating damage and interfaces with genome maintenance programs that influence mismatch repair processing and cell-cycle checkpoint signaling. Altered MGMT function or expression is widely studied in contexts of alkylation-induced genomic instability and tumor biology, where DNA repair capacity can shape mutation spectra and cellular stress responses. In mice, Mgmt provides a tractable system for investigating DNA damage response pathways, tissue-specific repair biology, and mechanisms linking alkylation lesions to carcinogenesis and neurotoxicity models.
MGMT CRISPR/Cas9 KO Plasmid (m) is a pool of plasmids designed for targeted disruption of the Mgmt gene in mouse cell lines. Each plasmid co-expresses a unique single guide RNA (sgRNA) targeting a distinct site within the Mgmt 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 Mgmt 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 MGMT protein expression.
This CRISPR knockout system enables efficient generation of Mgmt-deficient cell models for investigation of MGMT signaling, functional genomics studies, cancer biology research, and evaluation of therapeutic responses in human cell lines.
Key Features
- sgRNAs targeting Mgmt exon(s) critical for MGMT 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 Mgmt genomic sites to improve knockout efficiency
- Compatible with delivery by transfection
Design Variants
CRISPRs +/- HDRs
- gRNAs encoded by MGMT CRISPR/Cas9 KO Plasmid (m) and MGMT CRISPR/Cas9 KO Plasmid (m2) target distinct sites within the Mgmt locus. One or both targeting designs may be available. See Related Products for availability.
- HDR donor constructs encoded by MGMT HDR Plasmid (m) and MGMT HDR Plasmid (m2) contain a puromycin resistance cassette and an RFP reporter flanked by Mgmt homology arms to support homology-directed repair at defined Mgmt 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.